JP2022119840A - Crystalline polymorph of muscarinic acetylcholine receptor agonist - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide novel crystalline forms of a spiro-compound which acts as a muscarinic acetylcholine receptor agonist.

SOLUTION: The present invention relates to a crystalline polymorph of monohydrate Form III of the following compound having an X-ray powder diffraction pattern containing at least one of the following 2-theta values (±0.2): 12.3, 17.3, 17.5, 19.9, 21.6, 24.6, 26.3, and 35.4, as measured using CuKα radiation, and substantially free of peaks having 2 theta values in the range of 10.8-11.9.

SELECTED DRAWING: None

COPYRIGHT: (C)2022,JPO&INPIT

Description

The present invention relates to crystalline polymorphs of spiro compounds, pharmaceutical compositions containing such polymorphs, and their use in the treatment of acetylcholine-mediated diseases.

Degeneration of cholinergic neurons and cholinergic hypofunction are associated with Alzheimer's disease (A
D) related pathology. Muscarinic acetylcholine receptors (mAChRs) mediate acetylcholine-induced neurotransmission and five mAChR subtypes (M1-M5) have been identified. Among them, M1 receptors are widely expressed in the central nervous system (CNS) and have been implicated in many physiological and pathological brain functions. In addition, M1 receptors
D and several other neurodegenerative diseases are considered important therapeutic targets. The review is
For example, Jiang et al. , Neurosci. Bull. 30 (2014),
295-307. Xanomeline, an orthosteric muscarinic agonist with moderate selectivity for M1 and M4, was one of the first compounds to show improvement in behavioral disturbances in AD patients and efficacy in schizophrenia patients. rice field. On the other hand, attempts are being made to develop additional compounds that exhibit improved selectivity for the M1 receptor. A review can be found, for example, in Mel
Ancon et al. , Drug Discovery Today 18 (201
3), 1185-1199. However, in all clinical studies,
Xanomeline and other related muscarinic agonists are associated with nausea, gastrointestinal pain, and diarrhea.
rhhea], diaphoresis (excessive sweating), hypersalivation (excessive salivation), syncope, and bradycardia show an unacceptable margin of safety.

Therefore, it is possible to selectively modulate M1 receptor activity, does not exhibit the adverse effects seen with stimulation of other muscarinic receptors, is safe and tolerable in humans, and is currently manufactured. There remains a need for compounds that are compatible with drug manufacturing processes that comply with good control and quality control (cGMP) regulations.

This problem is solved by the invention according to the embodiments characterized in the claims and further described below.

The present invention generally relates to novel crystalline forms of (S)-2-ethyl-8-methyl-1-thia-4,8-diazaspiro[4.5]decan-3-one (Compound A).

ここで、上記多形は、
－ 図１に示されているパターンと実質的に同じ粉末Ｘ線回折（ＸＲＰＤ）パターン、
それぞれ図３Ａおよび３Ｂに示されている曲線と実質的に同じ示差走査熱量測定（ＤＳＣ
）曲線、図９Ｂに示されているスペクトルと実質的に同じ固相ＣＰ／ＭＡＳ ^１３Ｃ Ｎ
ＭＲスペクトル、ならびに図１０Ａに示されているスペクトルと実質的に同じＡＴＲ Ｆ
Ｔ－ＩＲスペクトルを示す、化合物Ａの一水和物である形態Ｉ；
－ それぞれ図６Ａおよび６Ｂに示されているパターンと実質的に同じＸＲＰＤパター
ン、それぞれ図７Ａおよび７Ｂに示されている曲線と実質的に同じ示差走査熱量測定（Ｄ
ＳＣ）曲線、それぞれ図８Ａおよび８Ｂに示されている曲線と実質的に同じ熱重量分析（
ＴＧＡ）曲線、図９Ａに示されているスペクトルと実質的に同じ固相ＣＰ／ＭＡＳ ^１３
Ｃ ＮＭＲスペクトル、ならびに図１０Ａおよび１０Ｂに示されているスペクトルと実質
的に同じＡＴＲ ＦＴ－ＩＲスペクトルを示す、化合物Ａの無水形態である形態ＩＩ；な
らびに
－ 図２に示されているパターンと実質的に同じＸＲＰＤパターン、図４Ａおよび４Ｂ
に示されている曲線と実質的に同じＤＳＣ曲線、図５に示されている曲線と実質的に同じ
ＴＧＡ曲線、ならびに図９Ｃに示されているスペクトルと実質的に同じ固相ＣＰ／ＭＡＳ
^１３Ｃ ＮＭＲスペクトルを示す、化合物Ａの一水和物である形態ＩＩＩ
からなる群から選択される。

where the above polymorph is
- a powder X-ray diffraction (XRPD) pattern substantially the same as the pattern shown in Figure 1,
Differential scanning calorimetry (DSC) curves substantially identical to the curves shown in FIGS. 3A and 3B, respectively.
) curve, solid-phase CP/MAS ¹³ CN substantially identical to the spectrum shown in FIG. 9B.
MR spectrum and ATR F, substantially the same as the spectrum shown in FIG. 10A
Form I, the monohydrate of Compound A, showing a T-IR spectrum;
- XRPD patterns substantially the same as those shown in Figures 6A and 6B, respectively, differential scanning calorimetry (D
SC) curves, substantially the same thermogravimetric (
TGA) curve, solid phase CP/MAS ¹³ substantially identical to the spectrum shown in FIG. 9A.
Form II, the anhydrous form of Compound A, which exhibits a C NMR spectrum and an ATR FT-IR spectrum substantially identical to those shown in FIGS. 10A and 10B; essentially the same XRPD patterns, FIGS. 4A and 4B
DSC curve substantially the same as shown in Figure 5, TGA curve substantially the same as the curve shown in Figure 5, and solid-phase CP/MAS curve substantially the same as the spectrum shown in Figure 9C.
Form III, the monohydrate of Compound A, showing a ¹³ C NMR spectrum
selected from the group consisting of

The crystalline polymorphic forms are useful for various pharmaceutical applications, for example, stimulation of M1 muscarinic receptors. The present invention relates to individual embodiments, illustrated in Figures 1 to 16 and specifically illustrated in the Examples, which demonstrate the essential characteristics of the crystalline polymorphic forms of the subject compound A of the present invention.

Compound A [(S)-2-ethyl-8-methyl-1-thia-4,8-diazaspiro [4.
5]Decan-3-one] is known to be a selective M1 muscarinic receptor agonist and has been identified in US Pat.
51 and 7,049,321 and corresponding International Application Publication No. 03/09258
0 listed.

AF267B increases αAPP, reduces Aβ levels and tau hyperphosphorylation, and inhibits Aβ-induced neurotoxicity in vitro through M1 receptor-mediated regulation of kinases such as PKC, MARK, and GSK3β. It has been shown to prevent Reviews can be found, for example, in Fisher, Curr. Alzheimer Res. 4 (2007)
, 577-580 and Fisher, J. et al. Neurochem. 120 (2012),
See 22-33. AF267B was found to improve spatial memory in 3xTg-AD mice and was associated with reduced Aβ and tau pathology in the hippocampus and cortex [Caccamo et al. , Neuron. 49 (2006), 671-6
82]. Compound A (AF267B), formulated as a drug designated NGX-267, has previously been shown in phase II clinical trials for the treatment of dry mouth and in phase I clinical trials for the treatment of cognitive impairment in Alzheimer's disease and schizophrenia. Phased clinical trials have also been performed. In this context, the actual manufacturing process for AF267B/NGX-267 used in clinical trials was not disclosed, but the laboratory-scale process described in the Examples of WO 03/092580 and the were thought to be different. However, despite promising clinical trial results in 2009, all clinical studies of this drug candidate were halted and never restarted.

Experiments performed according to the present invention have now surprisingly revealed that Compound A has three distinct crystalline polymorphs, two monohydrates of Compound A, referred to herein as Form I and Form III. , and one anhydrous Compound A, referred to herein as Form II, was found to exist. The three crystalline forms are sometimes referred to as polymorphs. Each of the three polymorphs is not a salt form. Anhydrous Form II and monohydrate Form III of Compound A were found to have activity substantially similar to that described for NGX-267 (“AF267B”). Since the intended use of this compound is as a therapeutically active drug, the most stable pharmaceutically acceptable form of the monohydrate of Compound A would be of great interest.

Therefore, a novel crystalline form, in particular the compound A[(S)-2-ethyl-8-methyl-1-thia-4,8-diazaspiro[4.5]decane-3-, which has favorable properties in pharmaceutical manufacture Anhydrous Form II and a monohydrate Form III of [on] are provided. Also methods for preparing the novel forms and for converting Form II to crystalline Form I or Form III and Form I or Form III to Form II, as well as M1 muscarinic receptor Methods are provided for preparing medicaments containing the novel crystalline forms suitable for use in treating diseases and disorders responsive to modulation of .

In one embodiment, the present invention encompasses crystalline polymorphic forms of Compound A that exhibit certain powder diffraction patterns. In one embodiment, the crystalline polymorphic form is substantially free of solvates. In preferred embodiments, such crystalline polymorphic forms are substantially free of water, ie, substantially anhydrous. In another embodiment, the crystalline polymorphic forms contain solvates. In preferred embodiments, the crystalline polymorphic form contains a certain amount of water.

In one embodiment, the present invention encompasses a method for producing crystalline polymorphic forms of Compound A that are solvate-free. In a preferred embodiment, the crystalline polymorphic form is free of water (
anhydrous).

In one embodiment, the present invention provides the following single crystal X-ray data: P2(1)a = 8.1416 (
13), (α=90°), b=7.9811(12) (β=90.761(2)°, c=
17.878(3), (γ=90°), Å, T=173(1) K
A solvent-free, water-free crystalline polymorphic form of Compound A (Form II) is provided. In one embodiment of the invention, the crystalline form has the following data: Volume = 1161.6(3) Å3, Z
= 4, F(000) = 464, theoretical density, Dc = 1.226 Mg/m ³ , absorption coefficient, μ =
It is further characterized by 0.251 mm ⁻¹ .

In another embodiment, the methods of the invention produce crystalline polymorphic forms of Compound A, including solvates. More preferably, such crystalline polymorphic forms contain water. Even more preferably, the crystalline polymorphic form is the monohydrate of Compound A.

In yet another embodiment, the invention encompasses methods of converting different crystalline polymorphs of the invention into each other. More preferably, the process converts the anhydrous crystalline polymorph and the monohydrate crystalline polymorph of the invention into each other.

More specifically, as noted above, the present invention provides three crystalline polymorphic forms of Compound A: Anhydrous Compound A, which for convenience will be referred to herein as Form II one of
Compound A containing four novel crystalline polymorphic forms; and one molecule of water for each molecule of Compound A.
provides two crystalline polymorphic forms (Form I and Form III) of

In addition, the present invention includes various methods for preparing crystalline Forms I, II, and III of Compound A.

In yet another set of embodiments, the invention encompasses methods and compositions for preparing and administering pharmaceutical compositions to treat mammalian diseases or conditions responsive to stimulation of M1 muscarinic receptors. , the method comprises providing a subject in need thereof with a crystalline polymorphic form of Compound A (and/or a pharmaceutically acceptable salt thereof) and a pharmaceutically acceptable carrier, diluent,
or administering an effective amount of the compound or mixture of compounds, including excipients, to stimulate M1 muscarinic receptors. In preferred embodiments, the crystalline polymorphic form is substantially free of water. In a more preferred embodiment, the crystalline form is Compound A Form II. In another preferred embodiment, the crystalline polymorphic form contains one molecule of water per molecule of Compound A,
Form III.

Yet another embodiment is a crystalline polymorphic form of Compound A disclosed herein, e.g.
. a crystalline polymorphic form exhibiting an X-ray powder diffraction pattern comprising at least one peak at a diffraction angle 2-theta selected from the group consisting of 9°, 10.8°, and 11.8° ± 0.2; A pharmaceutical composition comprising a pharmaceutically acceptable carrier, diluent, or excipient is included.

In one embodiment of the invention, the pharmaceutical compositions contemplated herein further comprise additional forms of Compound A in crystalline, solvate, or amorphous form. In certain embodiments, additional forms of Compound A are monohydrate forms. In another specific embodiment, the pharmaceutical composition comprises
at least 70% by weight of said crystalline polymorphic form, based on the total weight of Compound A in the composition;
It preferably contains 80%, 90%, 95%, or 99% by weight of the above crystalline polymorphic form.

Embodiments of the present invention are characterized by the following items, and according to the accompanying figures and examples:
will be explained in more detail.

FIG. 1 shows the X-ray powder diffraction patterns of Compound A crystalline Form I crystallized from ethyl acetate (A) and slowly evaporated from water (B). Figure 2 represents the X-ray powder diffraction pattern of the monohydrate crystalline polymorphic form of Compound A (Form III). Figure 3 shows differential scanning calorimetry (DSC) of the monohydrate crystalline polymorphic form of Compound A (Form I) crystallized from ethyl acetate (A) and slowly evaporated from water (B). ) represents the curve. DSC shows two endothermic peaks, one at about 107°C and the other at about 136.17°C. FIG. 4 depicts a differential scanning calorimetry (DSC) curve for the monohydrate crystalline polymorphic form of Compound A (Form III). A) From a reslurry in water. DSC shows two endothermic peaks, one at about 77.10°C and the other at about 134.87°C. B) cGMP active pharmaceutical ingredient (API) prepared by crystallization from acetone and 1.3 equivalents of water. DSC shows two endothermic peaks, one at about 61.18°C and the other at about 133.75°C. FIG. 5 depicts the results of thermogravimetric analysis (TGA) of the monohydrate crystalline polymorphic form of Compound A (Form III, API). When heated to temperatures up to about 110°C, the TGA shows a weight loss of 7.8%. Moisture by Karl Fischer analysis is 7.7% as expected from a stoichiometric monohydrate. 6 represents the X-ray powder diffraction pattern of anhydrous crystalline polymorphic Form II of Compound A. FIG. A) Recrystallization from acetone, B) cGMP API. FIG. 7 depicts the DSC curve for the anhydrous crystalline polymorphic Form II of Compound A. A) Recrystallization from acetone, B) cGMP API. DSC of A) and B) show only one endothermic peak at about 135.35° C. and 134.29° C. respectively. 8 depicts TGA results for anhydrous crystalline polymorphic Form II of Compound A. FIG. A) recrystallization from acetone (TGA showed no significant weight loss up to 110° C.), B) API (TGA showed 0.73% weight loss at 115° C.). FIG. 9 is a representative solid state CP/MAS ¹³ C NMR spectrum of crystalline Forms I and II of Compound A. Form III has the same CP/MAS ¹³ C NMR spectrum as Form I. Figure 10: A) Representative ATR FT-IR spectra of crystalline Form I (crystallized from ethyl acetate) and Form II (crystallized from dioxane) of Compound A. A discernible difference in some absorption peaks was observed. Each form is distinguished by the 1340-1390 cm ⁻¹ region of the spectrum, which region can be used to identify polymorphs. For crystalline form I, the relative shapes and intensities of the absorption peaks at 1352, 1369 and 1387 cm ⁻¹ can be used for diagnostic purposes. For crystalline form II, the relative shape and intensity of the absorption peak at 1340-1362 cm ⁻¹ can be used for identification purposes. B) cGMP API Crystalline Form II. FIG. 11 represents the single crystal structure of Compound A anhydrous crystals (Form II crystallized from acetonitrile) showing the molecules of two conformationally distinct asymmetric units and their arrangement in the unit cell. Figure 12 shows three X-ray powder diffraction patterns of compound A: A) simulation of single crystal Form II (see Figure 11) by using XPREP, B) experimental XRPD of crystalline material Form II (cGMP API). represents a pattern. FIG. 13 shows three X-ray powder diffraction patterns of Compound A: experimental XRPD patterns of Form I (crystallized from ethyl acetate), Form II (cGMP API), Form III (cGMP API) crystalline material, and XPREP. 1 represents a simulation of the single crystal morphology shown in US Pat. No. 7,439,251 B2 by using . FIG. 14 represents the single crystal structure of Compound A with one molecule of water (Form I) crystallized from ethyl acetate. Figure 15 represents the conversion of Form II to Form III at 95% RH. A) CP/MAS ¹³ C NMR solid-state NMR. Compound A anhydrous crystals: (A) dry and (B) after storage at room temperature for 1 week at 95% RH. B) Compound A anhydrous crystals were completely converted to pure monohydrate crystals, Form III, as indicated by CP/MAS ¹³ C solid-state NMR, DSC, and TGA. FIG. 16 depicts solid-state CP/MAS ¹³ C NMR of the conversion of Compound A crystalline Forms I/III to Form II as a function of time at 90°C. The arrow indicates the diagnostic peak of crystalline form II.

Definitions Throughout the specification and claims, terms defined when introduced retain their definitions throughout the specification and claims. In addition,
The following definitions apply throughout the specification and claims.

The terms "crystalline form" and "polymorph" refer to a particular chemical compound in a particular crystalline state, whether or not the chemical compound is solvated. Thus, for example, a chemical compound shown to crystallize in two different unsolvated forms and one solvated form is said to crystallize in three different crystalline polymorphic forms or polymorphs. become. Additionally, "polymorph" means a crystalline form of a substance that shares the same chemical formula, but which differs from another crystalline form.

With respect to "polymorphic purity", crystalline polymorphic Forms I-III of Compound A are preferably substantially free of chemical impurities (e.g., by-products produced during preparation of the polymorph) and other polymorphic crystalline forms. does not include For the purposes of the present invention, "substantially free" of chemical impurities means about 5 w/w%
less than or equal to chemical impurities, preferably less than or equal to about 3 w/w% chemical impurities, more preferably less than or equal to about 2 w/w% chemical impurities, and even more preferably less than about 1 w/w% or equivalent chemical impurities.
The terms "purified" or "in purified form" with respect to polymorphs are used herein after obtaining from one or more purification processes described herein or known to those of skill in the art. Refers to the physical state of said polymorphs characterized as sufficiently pure by standard analytical techniques described or well known to those skilled in the art. The purified forms of crystalline polymorphic Forms I-III of the monohydrate of Compound A are substantially free of chemical impurities.

The term "treatment or prevention" when used in reference to the disorders and conditions listed herein means amelioration, prevention or alleviation of the symptoms and/or effects associated with such disorders or conditions. The term "prevention," as used herein, refers to prophylactic administration of a medicament. To those skilled in the medical arts, the term "preventing" includes:
Understand that it is not an absolute term. In the medical arts, the term "prevent" refers to prophylactic administration of a drug to substantially reduce the likelihood or severity of a condition, which is the intended meaning of the claims. "Patient" includes both humans and other animals. "Mammal" includes humans and other mammals.

As used herein, "pharmaceutically acceptable" is physiologically tolerable when administered to humans and typically causes allergic reactions, such as stomach upset and dizziness. or refers to materials and compositions that do not cause similar adverse reactions. Typically, the term "pharmaceutically acceptable," as used herein, refers to any drug approved by a federal or state regulatory agency for use in animals, and more particularly in humans. or listed in the United States Pharmacopoeia or other generally recognized pharmacopoeia.

The phrase "pharmaceutically acceptable salt" as used herein includes, but is not limited to, acidic or basic groups that may be present in compounds used in the compositions. Contains salt. Compounds included in the present compositions that are basic in nature are capable of forming a wide variety of different salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds include non-toxic acid addition salts, namely, but not limited to, sulfuric acid, citric acid, maleic acid, acid, acetic acid, oxalic acid, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate [acid p
phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, oleate, tannate, pantothenate, hydrogen tartrate, ascorbic acid acid, succinate, maleate, gentisate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethane sulfonate, benzenesulfonate, p-
Form salts containing pharmacologically acceptable anions, including toluenesulfonate and pamoate [i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)] salts It is something to do. Compounds containing amino moieties included in the present compositions can form pharmaceutically acceptable salts with various amino acids, in addition to the acids mentioned above.

As used herein, the term "pharmaceutically acceptable carrier" can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules,
Included are cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances, which may also act as diluents, flavoring agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier is a finely divided solid that is in a mixture with the finely divided active ingredient. In tablets, the active ingredient or ingredients are mixed with a carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired. Liquid form preparations include solutions, suspensions, and emulsions, for example, water, water propylene glycol solution. For parenteral injection, liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.
Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents as desired. Aqueous suspensions suitable for oral use contain finely divided active ingredient in water with viscous materials, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well known suspending agents. can be produced by dispersing

As used herein, the terms "binder" or "excipient" refer to agents used to impart cohesive properties to powdered materials. Binders or "granulating agents" as they are sometimes known, impart cohesive properties to a tablet formulation, thereby ensuring that the tablet retains its integrity after compression. and enhanced free-flowing properties by formulating into granules of desired hardness and size. Materials commonly used as binders include starch; gelatin; sugars such as sucrose, glucose, dextrose, molasses, and lactose; mucilage of
isapol husks, carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone, Veegum, microcrystalline cellulose, microcrystalline dextrose, amylose, and natural or synthetic gums such as larch arabinogalactan. "Excipient" means an essentially inert substance used as a diluent or to give form or consistency to a formulation. In general, excipients are substances other than the active substance,
It can be defined as a component of a pharmaceutical form that is ingested by or administered to a patient. See, for example, Annex of Directive 2001/83/EC. Certain excipients can also serve as disintegrants, that is, to help disperse a solid pharmaceutical composition when exposed to body fluids.

As used herein, a "diluent" is an inert substance added to increase the bulk of the formulation and to compress tablets to a practical size. Commonly used diluents include calcium phosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride, dry starch, powdered sugar, silica and the like.

As used herein, "about,""approximately,""substantially," and "significantly" are understood by those of ordinary skill in the art and will vary to some extent with the context in which they are used. "About" and "approximately" shall mean plus or minus <10% of the specified term, where the use of the above terms would not be apparent to one of ordinary skill in the art given the context in which they are used; "
"Substantially" and "significantly" shall mean plus or minus >10% of the specified term.

When using NMR data to describe a material, the NMR peaks may be reported as absolute chemical shift values relative to a standard (e.g., tetramethylsilane), or alternatively, the material is the chemical shift of the peak. It will be appreciated that it may also be described as the difference between the values. Thus, for example, a material may be described as having an NMR spectrum with peaks at 170.0 and 130.0 ppm, or the material has a 40 ppm difference between the largest chemical shift value and another peak. It may be described as having an NMR spectrum that is 0.0 ppm. The latter reporting method is useful in that it is not affected by systematic errors that can occur in reporting absolute chemical shift values for materials. Also, NM
It will be appreciated that when using the R spectrum, the peak shape can assist in identifying the material. Thus, for example, two crystalline polymorphic forms of the same molecule may have NMR peaks at approximately the same chemical shift value, but in the spectrum of one of the materials the peaks are those of the other crystalline polymorphic form. It may be much narrower or much higher than the spectrum, thus facilitating identification of one or the other crystalline polymorphic form. The same applies mutatis mutandis to other analytical methods.

The above-characterized inventive subject matter and various aspects and embodiments thereof shall now be described with reference to the following figures and examples. Unless otherwise indicated,
The terms used are understood according to the definitions given herein.

Furthermore, the disclosure content of the above description in the background of the invention forms part of the disclosure of the present invention to the extent applicable. The present invention provides specific methodologies, protocols,
It should be understood that it is not limited to cell lines, excipients, carriers, and reagents per se. Also, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the invention. The scope of the invention shall be limited only by the appended claims. Embodiments of the present invention provide the following items [1] to [4
0]. Accordingly, in its broadest aspects, the invention relates to:
[1] Formula:

の化合物Ａの結晶性多形であって、
上記多形が、
一水和物形態Ｉであって、
（ｉ）ＣｕＫ_α線を使用して測定される以下の２シータ値（±０．２）：８．８、１
２．３、１７．５、１９．９、２１．６ ２３．５、２４．５、２６．３、２８．８、３
１．６の少なくとも１つを含有するが、以下の２シータ値、１７．３、１７．９、２１．
９、２４．９、２９．３、３０．８、および３３．４の少なくとも１つを欠くＸ線粉末回
折パターンを有し（図１も参照）、
（ｉｉ）結晶性形態のＺｎＳｅ ＡＴＲ－ＦＴ－ＩＲ吸収スペクトルが、１３５２、
１３６９、および１３８７ｃｍ^－１から選択される値を有する少なくとも１つの吸収ピー
クを含有し、かつ／または
（ｉｉｉ）結晶性形態が、示差走査熱量測定法（ＤＳＣ）により測定して、１０７．
１℃（１０４．８５℃から開始）および１３６．１７℃（１３３．４１℃から開始）に吸
熱ピークを示し（図３も参照）、任意選択で
（ｉｖ）結晶性形態の^１３Ｃ固相ＮＭＲが、ＴＭＳに対してｐｐｍ単位で表される以
下の化学シフト値：６７．０９、５４．０８、４６．５９、４０．９７、３０．１５、お
よび１３．２７の１つを有する少なくとも１つの共鳴を含有し、かつ／または
（ｖ）結晶性形態の^１３Ｃ固相ＮＭＲが、１０７．３、１２０．３、１２７．８、１
３３．４、１４４．２、もしくは１６１．１の、最も大きな化学シフトを有する共鳴と別
の共鳴との化学シフトの差を含有する、形態；
無水形態ＩＩであって、
（ｉ）以下の単結晶Ｘ線データ：Ｐ２（１）ａ＝８．１４１６（１３）、（α＝９０
°）、ｂ＝７．９８１１（１２）（β＝９０．７６１（２）°、ｃ＝１７．８７８（３）
、（γ＝９０°）、Å、Ｔ＝１７３（１）Ｋにより特徴付けられる単結晶Ｘ線を示し（図
１１も参照）、
（ｉｉ）ＣｕＫ_α線を使用して測定される以下の２シータ値（±０．２）：９．９、
１０．８、１１．８、１１．９、１４．８、１６．２、１８．２、１８．５、１９．８、
２１．３、２２．４、２３．９、２９．２、２９．７、および３３．１の少なくとも１つ
を含有するＸ線粉末回折パターンを有し（表１も参照）、
（ｉｉｉ）結晶性形態のＺｎＳｅ ＡＴＲ－ＦＴ－ＩＲ吸収スペクトルが、１９０６
、１３４０、１４４７、２８６９、２９０１、２９５１、および３００６～３０１２ｃｍ
^－１から選択される値を有する少なくとも１つの吸収ピークを含有し、
（ｉｖ）結晶性形態の^１３Ｃ固相ＮＭＲが、ＴＭＳに対してｐｐｍ単位で表される以
下の化学シフト値：１７５．０；６５．３、６４．０；４５．８、４５．０；４９．３、
４３．６、３９．５；３８．８；２８．９、２６．０；１５．４、１４．８の１つを有す
る少なくとも１つの共鳴を含有し（表５も参照）、
（ｖ）結晶性形態の^１３Ｃ固相ＮＭＲが、１０９．７もしくは１１１；１２９．２も
しくは１３０．０；１２２．７；１２５．７；１３１．４；１３５．５；１３６．２；１
４６．１もしくは１４９．０；および１５９．６もしくは１６０．２の、最も大きな化学
シフトを有する共鳴と別の共鳴との化学シフトの差を含有し、かつ／または
（ｖｉ）結晶性形態が、ＤＳＣにより測定して、１３４．２℃から開始し１３５．４
℃±０．２℃のピークを有する吸熱ピークを有し、１０６℃～１１０℃に吸熱ピークを実
質的に有しておらず、約５０℃～約１２０℃の範囲に吸熱ピークを欠く（図７も参照）、
形態；あるいは
一水和形態ＩＩＩであって、
（ｉ）ＣｕＫ_α線を使用して測定される以下の２シータ値（±０．２）：１２．３、
１７．３、１７．５、１９．９、２１．６、２４，４、２６，３、および３５．４の少な
くとも１つを含有し、１０．８～１１．９の範囲の２シータ値を有するピークを実質的に
含まないＸ線粉末回折パターンを有し（表２も参照）、
（ｉｉ）結晶性形態の^１３Ｃ固相ＮＭＲが、ＴＭＳに対してｐｐｍ単位表される以下
の化学シフト値：６７．５６、５４．６０、４７．９７、４１．４９、３０．７０、およ
び１３．７７の１つを有する少なくとも１つの共鳴を含有し（表５も参照）、
（ｉｉｉ）結晶性形態の^１３Ｃ固相ＮＭＲが、１０７．３、１２０．３、１２７．０
、１３３．４、１４４．２、もしくは１６１．１の、最も大きな化学シフトを有する共鳴
と別の共鳴との化学シフトの差を含有し、
（ｉｖ）結晶性形態のＺｎＳｅ ＡＴＲ－ＦＴ－ＩＲ吸収スペクトルが、１０３９、
１３５３、１３６９、１３６９、１３８８、２９１８、２９７４、および３０８８ｃｍ^－
１から選択される値を有する少なくとも１つの吸収ピークを含有し、かつ／または
（ｖ）結晶性形態が、ＤＳＣにより測定して、５８～９４℃に非常にブロードな吸熱
ピーク、および１３３．７℃から開始し１３４．９℃にピークを有する吸熱ピークを示す
（図４および表３も参照）、形態
からなる群から選択される、結晶性多形。

A crystalline polymorph of Compound A of
The above polymorph is
Monohydrate Form I,
(i) the following 2-theta values (±0.2) measured using CuK _α radiation: 8.8, 1
2.3, 17.5, 19.9, 21.6 23.5, 24.5, 26.3, 28.8, 3
1.6, but the following 2-theta values: 17.3, 17.9, 21.
having an X-ray powder diffraction pattern lacking at least one of 9, 24.9, 29.3, 30.8, and 33.4 (see also Figure 1);
(ii) the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1352;
1369, and 1387 cm ⁻¹ , and/or (iii) the crystalline form, as measured by differential scanning calorimetry (DSC), is 107.
showing endothermic peaks at 1° C. (starting at 104.85° C.) and ^136.17 ° C. (starting at 133.41° C.) (see also FIG. 3); has one of the following chemical shift values expressed in ppm relative to TMS: 67.09, 54.08, 46.59, 40.97, 30.15, and 13.27 and/or (v) the crystalline form has a ¹³ C solid-state NMR of 107.3, 120.3, 127.8, 1
forms containing the chemical shift difference between the resonance with the largest chemical shift and another resonance of 33.4, 144.2, or 161.1;
Anhydrous Form II,
(i) the following single crystal X-ray data: P2(1) a=8.1416(13), (α=90
°), b = 7.9811 (12) (β = 90.761 (2) °, c = 17.878 (3)
, (γ=90°), Å, T=173(1) K (see also FIG. 11) showing single crystal X-rays characterized by
(ii) the following 2-theta values (±0.2) measured using CuK _α radiation: 9.9;
10.8, 11.8, 11.9, 14.8, 16.2, 18.2, 18.5, 19.8,
having an X-ray powder diffraction pattern containing at least one of 21.3, 22.4, 23.9, 29.2, 29.7, and 33.1 (see also Table 1);
(iii) the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1906
, 1340, 1447, 2869, 2901, 2951, and 3006-3012 cm
containing at least one absorption peak having a value selected from ^-1 ;
(iv) ¹³ C solid state NMR of the crystalline form with the following chemical shift values expressed in ppm relative to TMS: 175.0; 65.3, 64.0; 45.8, 45.0; 49.3,
43.6, 39.5; 38.8; 28.9, 26.0; 15.4, 14.8 (see also Table 5);
(v) the crystalline form has a ¹³ C solid-state NMR of 109.7 or 111; 129.2 or 130.0; 122.7; 125.7; 131.4;
46.1 or 149.0; and 159.6 or 160.2, and/or Starting at 134.2°C and 135.4°C as measured by DSC
It has an endothermic peak with a peak at ±0.2°C, has substantially no endothermic peak at 106°C to 110°C, and lacks an endothermic peak in the range from about 50°C to about 120°C (Fig. 7),
form; or monohydrate Form III, wherein
(i) the following 2-theta values (±0.2) measured using CuK _α radiation: 12.3;
containing at least one of 17.3, 17.5, 19.9, 21.6, 24,4, 26,3, and 35.4 and having a 2-theta value ranging from 10.8 to 11.9 having an X-ray powder diffraction pattern (see also Table 2) that is substantially free of peaks with
(ii) ¹³ C solid-state NMR of the crystalline form, expressed in ppm relative to TMS, with the following chemical shift values: 67.56, 54.60, 47.97, 41.49, 30.70, and contains at least one resonance with one of 13.77 (see also Table 5);
(iii) the crystalline form has a ¹³ C solid state NMR of 107.3, 120.3, 127.0;
, 133.4, 144.2, or 161.1, containing the chemical shift difference between the resonance with the largest chemical shift and another resonance;
(iv) the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1039;
1353, 1369, 1369, 1388, 2918, 2974, and 3088 cm ⁻
contains at least one absorption peak having a value selected from ¹ , and/or (v) the crystalline form has a very broad endothermic peak at 58-94° C. and 133.7 as measured by DSC; A crystalline polymorph selected from the group consisting of forms exhibiting an endothermic peak with an onset at 0 C and a peak at 134.9 0 C (see also Figure 4 and Table 3).

Compound A is (2S)-2-ethyl-8-methyl-1-thia-4,8-diazaspiro [
4.5] International Application Publication No. 03/09258 as Decane-3-one (“AF267B”)
0A2 and reflects the following chemical structure.

This reference also describes chiral separation of the R and S forms of compound A by elution with an 85:15 mixture of acetonitrile/ethanol, followed by addition of more ethanol and evaporation to dryness to remove residual acetonitrile. mentioned to be removed. However, the methods for preparing Compound A disclosed in WO 03/092580 and related patents are not controlled with respect to specific polymorphic crystalline forms and thus
Previously, the polymorphic forms of Compound A were unknown. Furthermore, International Application Publication No. WO 03/09258
Compound A, prepared according to the method disclosed in No. 0, has not proven suitable as a drug for pharmaceutical compositions. Accordingly, in one embodiment, the present invention is based on U.S. Pat.
, 439,251 and 7,049,321 and corresponding International Application Publication No. 03
092580 and its preparation method.

As mentioned, an essential part of the present invention are three separate crystalline polymorphs of Compound A, two monohydrates of Compound A, referred to herein as Forms I and III, and this Based on the unexpected observation that one anhydrous Compound A, referred to herein as Form II, was found to exist.

It is known that active pharmaceutical ingredients often occur in different crystalline forms, or polymorphic forms that can be attributed to the lack of a crystalline state. As a result of the different ordering of the molecules in the crystal lattice, such polymorphic forms of the compounds differ in their melting points, X-ray diffraction patterns, infrared absorption fingerprints, and solid-state NMR spectra. therefore,
These are solids that share the same molecular formula but differ in the quantifiable features commonly used to characterize solids.

Polymorphism can also affect physical parameters that are important in pharmaceutical formulation and drug product manufacturing, such as storage stability, compressibility, and density. For example, 1
One form may be more likely to form desirable or undesired solvates, or may contain impurities, due to differences in the particle shape and size distribution of one form compared to another form. It can be difficult to filter and wash away. Polymorphism also affects the absorption, pharmacokinetics,
and/or have a direct pharmacological effect by affecting the rate of dissolution in physiological fluids, which may alter bioavailability. Ultimately, even the required dosage content in drug formulations required to obtain ideal drug efficacy may be different for different polymorphs. If the drug crystallizes as two or more crystalline forms with different bioavailability, the optimal dose will depend on the crystalline forms present in the formulation. Polymorphism can also affect pharmaceutical parameters that are important in formulation and product manufacturing, such as storage stability, compressibility, and density. One form may be more likely to form desirable or undesired solvates, or contain impurities, due to differences in particle shape and size distribution of one form compared to another form. It can be difficult to filter and wash free.

The present invention provides a crystalline form, namely the compound (S)-2-ethyl-8-methyl-1-thia-4.
,8-diazaspiro[4.5]decan-3-one (Compound A), processes for preparing them, and uses thereof. According to an embodiment of the present invention, there are three crystalline forms of Compound A: one crystalline form of anhydrous Compound A, which for convenience will be referred to herein as Form II; Two compounds of Compound A containing one molecule of water for each molecule of Compound A, which will be referred to herein as Forms I and III, respectively.
Two distinct crystalline forms are provided.

According to the present invention, the crystalline forms of Compound A can be characterized in various ways. For example, Form II can be readily separated from Forms I and III in the presence or absence of water using techniques such as those known in the state of the art, such as differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA), among others. can be distinguished. The morphology also includes, but is not limited to, the following techniques: X
X-ray powder diffraction (XRPD), solid-state carbon-13 nuclear magnetic resonance ( ¹³ C CP-MAS NMR) using cross-polarization magic angle spinning, attenuated total reflection Fourier transform infrared spectroscopy (ATR)
-FT-IR), DSC, and TGA, facilitated by a physical parameter or group of physical parameters that are present in one form but not the other. can be distinguished.

In one embodiment of the invention, the polymorph is monohydrate Form I, characterized in item [1] above and shown in Figures 3, 9, and 10, respectively.

Form I is preferentially obtained by direct crystallization of compound A from water-miscible organic solvents containing traces of water (ethanol, ethyl acetate, isopropanol, tert-butyl methyl ether, tetrahydrofuran) or water. It is a monohydrate form (Examples 1-6) and DSC shows two endothermic peaks, one at about 107° C. and the other at 136.17° C. (FIG. 3). Form I is a crystalline polymorph characterized by its XRPD peaks (Figure 1). Form I can be converted to anhydrous Form II by heating the material to 90° C. (Example 17). Form I can be converted to anhydrous Form II by heating the material to 160° C. and allowing the molten mass to crystallize at room temperature (Example 18).

In certain preferred embodiments, the present invention relates to crystalline polymorphic forms of Compound A that have proven particularly suitable for cGMP production. That is, the present invention relates to the following.
[2] The crystalline polymorph of [1], which is polymorph II.

According to an embodiment of the present invention, the following single crystal X-ray data: P2(1)a = 8.1416 (
13), (α=90°), b=7.9811(12) (β=90.761(2)°, c=
17.878(3), (γ=90°), Å, T=173(1) K
A crystalline polymorphic form of Compound A (hereinafter referred to as "Form II") is provided that is substantially free of water. In an embodiment of the present invention, the crystalline form has the following data: Volume = 1161.6
(3) Å3, Z=4, F(000)=464, theoretical density, Dc=1.226 Mg/m ³ ,
Further characterized by the absorption coefficient, μ=0.251 mm ⁻¹ (Example 26, FIG. 11)
.

Anhydrous crystalline Form II can be obtained by directly recrystallizing Compound A from a solvent such as acetonitrile, acetone, hexane, dioxane, cyclohexane, diethyl ether, preferably any solvent free of water, to form a suspension of Compound A in hexane. Obtained by phase equilibration (Examples 7-13). Form II is the crystalline polymorph shown by its XRPD (FIG. 6, respectively).
A and FIG. 6B). Various solvents can be used to produce Form II in good recovery. However, many of these solvents were not suitable for large scale due to the fact that the final product, long needles, stuck to the sides of the flask. Manual scraping was required to remove these sticky solids. Acetone was found to be one solvent that produced a white material with minimal loss of solids to the sides of the flask. Form II is reproducible with high purity and yield (88-93%) from acetone in a process that is scalable to the kilogram range and validated according to current manufacturing and quality control (cGMP) practices. It can be precipitated and crystallized well. Anhydrous crystalline Form II is stable under dry conditions, but upon exposure to ambient humidity for 3 months at room temperature, it converts to monohydrate Form I (Example 20a), increasing 95% over 1 week. % relative humidity converts to Form III (Example 15). API Form II converted to Form I after exposure to 90% relative humidity for 3 hours (Example 20b), and upon exposure to ambient humidity for 4 months at room temperature, Form I and Form III (Example 20c). API Form II can be obtained with a high degree of reproducibility and consistency when crystallized and dried according to well defined and controlled conditions. API Form II is highly stable under dry storage conditions for at least two years and has been tested in preclinical studies and clinical trials. In further embodiments, the invention relates to:
[4] The following 2-theta values measured using CuK _α radiation: 9.9, 10.8, 11.
8, 11.9, 14.8, 16.2, 18.2, 18.5, 19, 8, 21.3, 22.
at least 2, 3, 4, 5, 6, 7, 8, 9, 10 of 4, 23.9, 29.2 and 29.7, 33.0, 33.1 , 11, or all of the crystalline polymorphic Form II of [2] or [3].

According to the present invention, crystalline polymorphs of Compound A can be characterized in various ways. For example, in one embodiment, the crystalline polymorphs of Compound A are 9.9°, 10.8°, and 11.5°.
XR containing at least one peak at a diffraction angle 2θ selected from the group consisting of 8°±0.2
PD pattern is shown. In a related embodiment, the crystalline form of Compound A exhibits an XRPD pattern comprising 2-theta values of 9.9° and 10.8°. In another related embodiment, the crystalline form of Compound A exhibits an XRPD pattern comprising 2-theta values of 9.9° and 11.8°.
In yet another related embodiment, the crystalline polymorphs of Compound A are 10.8° and 11.8°
FIG. 3 shows the XRPD pattern with 2θ values in degrees. In yet another related embodiment, the crystalline form of Compound A exhibits an XRPD pattern comprising 2-theta values of 9.9°, 10.8°, and 11.8°.

In one embodiment of the invention, the crystalline form of Compound A has an X-ray powder diffraction pattern of 14.8
and 19.8°±0.2 at a diffraction angle 2θ selected from the group. In a related embodiment, the X-ray powder diffraction pattern of the crystalline form of Compound A further exhibits additional peaks at diffraction angles 2-theta of both 14.8° and 19.8°±0.2.

In another embodiment of the invention, the XRPD pattern of the crystalline form of Compound A is 18.2°
and at least one additional peak at a diffraction angle 2θ selected from the group consisting of: and 18.5°±0.2. In a related embodiment, the XRPD pattern of the crystalline polymorph of Compound A further comprises additional peaks at diffraction angles 2-theta of both 18.2° and 18.5°±0.2. Accordingly, in one embodiment, the invention relates to:
[5] The diffraction angle 2θ whose X-ray powder diffraction pattern is selected from the group consisting of 14.8° and 19.8° and/or selected from the group consisting of 18.2° and 18.5°
The crystalline polymorph Form II of any one of [2]-[4], comprising at least one additional peak in

In still further embodiments, the present invention relates to:
[6] The ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 906, 134
containing at least two, three, four, five, six, or all absorption peaks having values selected from 0, 1447, 2869, 2901, 2951, and 3006-3012 cm ⁻¹ , [2 ] to the crystalline polymorphic Form II of any one of [5].

[7] ¹³ C solid state NMR of the crystalline form with the following chemical shift values expressed in ppm relative to TMS: 175.0; 65.3, 64.0; 45.8, 45.0; 49.3, 44
. 28.9, 26.0; 15.4, 14.8 with at least 2, 3, 4, 5, 6, 7, or all resonances containing [2] to [6
].

In a further embodiment of the invention, ¹³ C solid phase NM of the crystalline polymorphic form of Compound A of the invention.
The R spectra are approximately 64.0, 45.0, 38.8, and 26.0±0.2 ppm
at least one peak having a chemical shift selected from the group consisting of In a related embodiment, the crystalline polymorphic Form II of Compound A has approximately 64.0 and 45.0±0.
Contains a peak with a chemical shift of 2 ppm. In another related embodiment, crystalline polymorphic Form II of Compound A comprises peaks with chemical shifts of approximately 64.0 and 38.8±0.2 ppm. In yet another related embodiment, the crystalline polymorphic Form II of Compound A is
Contains peaks with chemical shifts of approximately 64.0 and 26.0±0.2 ppm. In yet another related embodiment, the crystalline polymorphic Form II of Compound A is approximately 45.0 and 3
It contains a peak with a chemical shift of 8.8±0.2 ppm. In a further related embodiment, crystalline polymorphic Form II of Compound A comprises peaks with chemical shifts of approximately 45.0 and 26.0±0.2 ppm. In yet another related embodiment, crystalline polymorphic Form II of Compound A comprises peaks with chemical shifts of approximately 38.8 and 26.0±0.2 ppm.

In yet another related embodiment, the crystalline polymorphic Form II of Compound A is approximately 64.0,
45.0, and peaks with chemical shifts of 38.8±0.2 ppm. In yet another related embodiment, the crystalline polymorphic Form II of Compound A is approximately 64.0, 45.0
, and a peak with a chemical shift of 26.0±0.2 ppm. In further related embodiments, the crystalline polymorphic Form II of Compound A is approximately 64.0, 38.8, and 2
Contains peaks with chemical shifts of 6.0±0.2 ppm. In yet another related embodiment, crystalline polymorphic Form II of Compound A is approximately 45.0, 38.8, and 26.0 ±
Contains a peak with a chemical shift of 0.2 ppm. In yet another related embodiment, the crystalline polymorphic Form II of Compound A is approximately 64.0, 45.0, 38.8, and 26.0±
Contains a peak with a chemical shift of 0.2 ppm. In certain preferred embodiments, the invention relates to:
[8] having an X-ray powder diffraction pattern containing the following 2-theta values: 9.9, 10.8, 18.5, 19.8 ± 0.2, and a ¹³ C solid-state NMR of the crystalline form , ppm relative to TMS
The following chemical shift values expressed in units: 14.8, 15.4, 26.0, 28.9, 64.
ZnSe ATR-FT-IR absorption spectrum of the crystalline form contains an absorption peak with a value selected from the range of 1340-1362 cm ⁻¹ ±5 cm ⁻¹ , Crystalline polymorph Form II according to any one of [2] to [7].

[9] an X-ray powder diffraction pattern substantially as shown in FIG. 6, a solid-state 13 C NMR spectrum substantially as shown in FIG. 9, and a solid-state ¹³ C NMR spectrum substantially as shown in FIG. Like A
Crystalline polymorph Form II according to any one of [2] to [8], having at least one of a TR-FT-IR spectrum and a DSC pattern substantially as shown in FIG.
.

In another embodiment, the differential scanning calorimetry thermogram of the crystalline polymorphic form of Compound A of the present invention has an endothermic peak in the range of about 133°C to about 136°C, but an endothermic peak in the range of about 50°C to about 110°C. Lacks a range of endothermic peaks. See also FIG. In certain embodiments, the crystalline form further exhibits less than 1% weight loss when subjected to heating from about room temperature up to about 110° C. at a heating rate of about 3° C. per minute [thermogravimetric analysis (TGA)]. See also FIG.
Accordingly, in one embodiment, the invention relates to:
[10] Any one of [2] to [9], exhibiting less than 1% weight loss as determined by thermogravimetric analysis (TGA) at a heating rate of about 3°C per minute to a temperature of about 110°C. crystalline polymorphic form II as described in .

[11] The crystalline polymorph of [1], which is polymorphic Form III.

According to a preferred embodiment of the present invention, there is provided a crystalline monohydrate form of Compound A (hereinafter referred to as "Form III"). Form III is XRPD, ¹³ C, as described above.
Although the P/MAS ¹³ C NMR, ATR-FT-IR patterns are similar to Form I, its DSC pattern is quite different from Form I, with Form III having a very shallow and broad water spectrum in the range of about 60-80°C. loss and does not show the DSC/TCA peak at 104° C. characteristic of Form I (see FIG. 4 vs. FIG. 3). Form I and Form III are two different polymorphs shown by DSC/TGA. Without wishing to be bound by theory, we believe that the crystal matrix of Forms I and III is identical, but in Form I crystals the water is hydrogen bonded (see FIG. 14). ), whereas in Form III, water is presumably physically absorbed into the crystal pores, leaving the specific water-binding sites of the crystal substantially unoccupied.

Monohydrate Form III can be scaled up to the kilogram range and is process validated according to current manufacturing practices and quality control (cGMP) without any impurities of Form II or Form I and acetone can be reproducibly precipitated and crystallized in high yield (92%) from a solution of compound A in a mixture of and 1.3 equivalents of water. Example 14. Similar to Form II, Form III is less sticky than Form I and thus easier to filter, scrape, and handle than previously known solid compound A. Form III is stable for at least two years under various storage conditions and has been tested in preclinical studies and clinical trials.

Monohydrate Form III could not be produced by exposing monohydrate Form I to 95% relative humidity (Example 21), and such treatment provided more than one per molecule of Compound A. Nor does it produce another crystalline form containing many water molecules. Accordingly, in this aspect, the invention particularly relates to:
[12] The following 2-theta values measured using CuK _alpha radiation: 8.8, 12.3, 17
. The crystalline polyacrylamide according to [11] having an X-ray powder diffraction pattern containing at least two, three or all of 3, 17.5, 17.8 and 23.0 (see also Table 2) Form Form III.

[13] The X-ray powder diffraction pattern has the following 2-theta values measured using CuK _α radiation: 12.3, 19.9, 21.6, 24.6, 26.3, 31.6 , and 35.4
(see also Table 2).

[14] The crystal according to any one of [12] to [13], wherein the X-ray powder diffraction pattern does not substantially contain peaks having 2-theta values in the range of 10.8 to 11.9. sexual polymorphic form III.

[15] The following 2-theta values measured using CuK _alpha radiation: 12.2, 17.3, 1
X containing at least one of 9.9, 21.6, 24.6, 26.3, and 31.6
having a ray powder diffraction pattern, wherein said X-ray powder diffraction is substantially free of peaks having 2-theta values in the range of 10.8-11.9 (see also FIG. 2), [11]-[14 3. Crystalline polymorphic Form III according to any one of the claims

[16] The X-ray powder diffraction pattern has the following 2-theta values measured using CuK _α radiation: 12.2, 17.3, 17.5, 19.9, 21.6, 24.6 , 26.3.31
. 2, and at least 2, 3, 4, 5, or all of 35.4;
Crystalline polymorphic Form III of any one of [11] to [15].

[17] ¹³ C solid state NMR of the crystalline form with the following chemical shift values expressed in ppm relative to TMS: 67.56, 54.60, 47.07, 41.49, 30.70, and containing at least two, three, four, or all resonances of 13.77, [
11] to [16], the crystalline polymorphic form III.

[18] C solid-state NMR of the crystalline form is 107.3, 120.3, 127.9, ¹
contains at least 2, 3, 4, or all chemical shift differences between the resonance with the largest chemical shift and other resonances selected from 33.4, 144.2, and 161.1 , crystalline polymorphic form III of any one of [11] to [17].

[19] The ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1039,1
353, 1369 1369, 1388, 2918, 2974, and 3088 cm ^-1
The crystalline polyacrylamide according to any one of [11] to [18], containing at least 2, 3, 4, 5, 6, or all absorption peaks having a value selected from form form III
.

[20] The following 2-theta values 12.3, 17.3, 17.5, 19.9, 21.6±0.
2 and the ¹³ C solid state NMR of the crystalline form has the following chemical shift values expressed in ppm relative to TMS: 13.77, 30.70, 67.7. 56
The ZnSe ATR-FT-IR absorption spectrum of the crystalline form contains a resonance with
containing absorption peaks with values selected from 1353, 1369, and 1388±5 cm ⁻¹ , the crystalline form exhibits a very broad endotherm between 58-94° C. and an endotherm beginning at 133.9° C. A crystalline monohydrate form of the compound (S)-2-ethyl-8-methyl-1-thia-4,8-diazaspiro[4.5]decan-3-one.

[21] The crystalline polymorphic form of any one of [11]-[18] containing one molecule of water per molecule of Compound A, as shown in FIGS. III.

Another embodiment of the invention encompasses crystalline polymorphs of Compound A that are substantially free of solvates. In currently preferred embodiments, the crystalline form is substantially free of water.

In yet another embodiment of the invention, the crystalline polymorphic form of Compound A is 9.9°, 10.8°
, and 11.8°±0.2, wherein the morphology is substantially free of water. In certain embodiments, the forms contain less than about 2% water by weight. In further embodiments, the invention relates to:
[22] I-III according to any one of [1]-[21], which at all times is in substantially pure form, i.e. free of impurities and consists essentially of only one polymorphic form. Any one crystalline polymorphic form.

Also according to embodiments of the present invention are methods for preparing crystalline Forms I, II, and III of Compound A, methods for converting Form I to Form II and vice versa, and Form Methods are provided for converting Form II to Form III. Also provided in accordance with embodiments of the present invention are pharmaceutical compositions comprising one or more of crystalline Forms II or III of Compound A and a pharmaceutically acceptable carrier. Accordingly, in further aspects, the invention relates to the following embodiments.
[23] Compound A according to any one of [1] to [22]:

の結晶性多形形態を調製するためのプロセスであって、
（ａ）化合物Ａを適切な溶媒に溶解するステップ、
（ｂ）必要に応じて、得られた溶液を冷却するステップ、
（ｃ）形態ＩＩ結晶が析出するまで、十分な時間待機して、結晶性形態を結晶化させ
るステップ、および
（ｄ）上記結晶性形態を濾過するステップ
を含むプロセス。

A process for preparing a crystalline polymorphic form of
(a) dissolving Compound A in a suitable solvent;
(b) optionally cooling the resulting solution;
(c) waiting a sufficient time to crystallize the crystalline form until Form II crystals precipitate; and (d) filtering said crystalline form.

[24] The crystalline form is polymorphic Form II of any one of [2] to [10], and the solvent is acetone, acetonitrile, cyclohexane, hexane, dioxane, and a mixed solvent of ethanol and acetonitrile. The process of [23], selected from the group consisting of:

[25] The crystalline form is polymorphic Form III of any one of [11] to [20], by adding 1.3 moles of deionized water to an acetone solution of Compound A. resulting in the process of [23].

[26] The process of [25], wherein polymorphic Form III is obtained by reslurrying Compound A and/or polymorphic Form II in deionized water and filtering.

[27] The crystalline form is polymorphic form I according to [1], a water-miscible organic solvent containing trace amounts of water (ethanol, ethyl acetate, isopropanol, tert-butyl methyl ether, tetrahydrofuran), water or by slow evaporation of a solution of the compound dissolved in either water or ethyl acetate, [
23].

[28] the crystalline form is a mixture of crystalline polymorphs I and II, and the solvent is toluene;
selected from the group consisting of dichloromethane, 1-butanol, or diethyl ether;
The process of [23].

[29] A process for converting polymorphic form II to polymorphic form III, comprising: [2]
to the crystalline polymorphic Form II of any one of [10] at room temperature and at least 95%
at a relative humidity of for a time sufficient for conversion to crystalline polymorphic Form III according to any one of [11] to [20] (see also Figure 15).

[30] A process for converting polymorphic form I to polymorphic form II of any one of [2]-[10], comprising:
(a) maintaining crystalline Form I at an elevated temperature below the melting point of the crystalline form for a sufficient time to convert the crystalline form to said crystalline polymorphic Form II;
(b) suspending crystalline polymorphic Form I in a solvent selected from the group consisting of acetonitrile, cyclohexane, hexane, dioxane, and mixed solvents of ethanol and acetonitrile, and waiting for a sufficient time to obtain [2]- crystallizing the crystalline form of any one of [10] and filtering said crystalline form; and (c) heating crystalline polymorphic Form I above its melting point. forming a molten mass and cooling the molten mass.

The present invention relates to single crystal X-ray, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC),
Thermogravimetric analysis (TGA), and solid phase cross-polarization magic angle spinning nuclear magnetic resonance spectroscopy (CP/
MAS ¹³ C NMR), ATR FT-IR, etc., as well as those known in the art, as well as analytical techniques to be developed in the future, particularly due to the presence or absence of solvates. It includes several crystalline polymorphic forms of Compound A that can be distinguished. Accordingly, in one embodiment, the invention relates to:
[31] The resulting crystalline polymorphic Form I, II, or III is measured by its X-ray powder diffraction pattern in 2-theta values using CuK _alpha radiation, ZnSe ATR-FT-IR absorption spectrum by DSC. Measured endothermic peak, TGA, and/or ¹³ C solid-phase NM
[23]-[30], selected by specifying the polymorphism by the resonance measured by R
].

[32] A process for stably maintaining the crystalline polymorphic Form II of any one of [2] to [10], wherein said crystals are maintained at room temperature under a dry atmosphere. process that involves

Another embodiment of the invention is a pharmaceutical composition comprising a crystalline polymorphic form of Compound A substantially free of solvates and a pharmaceutically acceptable carrier, diluent or excipient therefor encompasses In some embodiments, the pharmaceutical compositions contemplated herein further comprise additional forms of Compound A in crystalline, solvate, or amorphous form. Accordingly, in further embodiments, the invention relates to:
[33] A pharmaceutical composition comprising the crystalline polymorph of any one of [1] to [22] and at least one pharmaceutically acceptable excipient or carrier.

In certain embodiments, the crystalline polymorphic form of Compound A is an anhydrous form. In another particular embodiment, the pharmaceutical composition comprises at least 70% by weight of crystalline Form II, preferably 80%, 90%, 95% by weight, based on the total weight of Compound A in the composition. or 99% by weight of the crystalline form. In certain embodiments, additional monohydrate polymorphic forms of Compound A exist. In another particularly preferred embodiment, the pharmaceutical composition comprises at least 70% by weight of crystalline Form III, preferably 80%, 90% by weight, based on the total weight of Compound A in the composition.
, 95%, or 99% by weight of the crystalline form.

A pharmaceutical composition according to the invention intended for parenteral administration comprises a crystalline polymorphic form of Compound A for dissolution or suspension in an aqueous or non-aqueous medium suitable for use in sterile injectable or infusible solutions. You can stay. Such pharmaceutical compositions containing Compound A polymorphic forms, or the vehicles in which they are intended to be dissolved or suspended, are intended to contain pharmaceutically acceptable antioxidants, buffers, and ready-to-use formulations. It may also contain a compound that renders it substantially isotonic with the blood of the recipient. Pharmaceutical compositions may also contain preservatives such as, for example, bacteriostatic or antimicrobial compounds; solubilizers, stabilizers, colorants, and odorants. Pharmaceutical compositions may also contain one or more adjuvants and/or therapeutically active agents in addition to the crystalline polymorphic forms of the present invention. The compositions may be presented in unit-dose or multi-dose containers, such as sealed ampoules and vials. In further embodiments, the invention relates to:
[34] polymorphic Form II or Form III from 1 mg to 100 mg, preferably 10 mg
The pharmaceutical composition according to [33], present in the formulation in an amount of ~50 mg, preferably the formulation is granular.

Pharmaceutical compositions intended for oral administration may be presented as discrete units such as capsules or tablets, or may be presented as powders or granules. Suitable excipients for tablets or hard gelatin capsules include lactose, corn starch or derivatives thereof, stearic acid or salts thereof. Excipients suitable for use in soft gelatin capsules include, for example, vegetable oils, waxes, fats, semisolid or liquid polyols, and the like.

In one embodiment of the invention, oral tablets can be prepared by direct compression molding of Compound A anhydrous crystalline Form II. In general, the advantages of direct compression molding are known to include fewer manufacturing steps involved, physical stability, and exclusion of heat and moisture. Direct compression tablets according to the present invention may additionally contain binders, disintegrants, and coloring agents such as those well known to those skilled in the art. In another embodiment, a pre-manufactured oral capsule is compound A crystalline Form I with excipients.
Contains II. After tablet compression or capsule closure, such preparations of the present invention may be coated with pharmaceutically acceptable coatings to further modify the release characteristics of the active agent in the gastrointestinal tract. good. The selection of the optimal site of release depends on the type of disease, the intended peak plasma concentration, the intended plasma time/concentration profile, and the intended time/concentration profile at the target site. In further embodiments, the invention relates to:
[35] A process for preparing a medicament suitable for oral administration based on the formulation of crystalline polymorph Form II of any one of [2] to [10], wherein the formulation is transformed into a tablet and direct compression molding process.

[36] A process for preparing a medicament suitable for oral administration based on the formulation of crystalline polymorphic Form III of any one of [11]-[22], comprising one or mixed with multiple excipients (pregelatinized starch, microcrystalline cellulose, colloidal silicon dioxide, and stearic acid) such that the mixture provides 5 mg or 10 mg of polymorph Form III per capsule; The process of filling size 4 white opaque hard gelatin halves capsules that can be used as an oral formulation for immediate release in the gastrointestinal tract.

According to the present invention, the daily dose of Compound A crystalline form may vary between 1 mg and 100 mg. In preferred embodiments of the invention, the daily dose may vary between 5 mg and 80 mg. In more preferred embodiments, the daily dose may vary between 10 mg and 50 mg. The exact amount of a single dose, frequency and schedule of compound administration, and duration of treatment will depend on the age, condition, and size of the subject and the severity of the condition being treated,
and observed undesired effects of treatment, and will be determined according to the judgment of the attending physician.

Another embodiment of the present invention is the modification of M1 muscarinic acetylcholine receptors such that diseases or conditions associated with cholinergic dysfunction (specifically understimulation of M1 receptors) are treated, ameliorated, or prevented. Any crystalline polymorphic form of Compound A, or such crystalline polymorphic forms and pharmaceutically acceptable It includes the use of pharmaceutical compositions containing salts of Accordingly, in further embodiments, the invention relates to:
[37] preferably at daily doses of about 10 mg and 50 mg for use in the treatment of medical conditions responsive to treatment, amelioration, or prophylaxis with muscarinic receptor agonists
The crystalline polymorphic form of any one of [1] to [22], wherein

[38] The conditions are diseases or conditions associated with cholinergic dysfunction, diseases or conditions in which cholinergic function is imbalanced, diseases or conditions associated with impaired activity of acetylcholine receptors, and activity of M1 receptors A crystalline polymorphic form for use according to [37], including a disease or condition associated with a disorder. Such diseases and conditions include, but are not limited to, Alzheimer's disease-type senile dementia; Alzheimer's disease (AD); Lewy body dementia combined with Alzheimer's disease and Parkinson's disease; Parkinson's disease; atrophy;
Multi-infarct dementia (MID), frontotemporal dementia; vascular dementia; stroke/ischemia, stroke/
Combined MID and AD; human head trauma; traumatic brain injury; age-related memory impairment; transient global amnesia syndrome; mild cognitive impairment (MCI); MCI;
Cognitive impairment (including amnesia, acute confusional disorder, attention deficit disorder, and concentration disturbance); hallucinatory delusional states, emotional attention disturbance; sleep disturbance; postoperative delirium; adverse effects of tricyclic antidepressants, schizophrenia Adverse Effects of Certain Drugs Used to Treat Parkinson's Disease and Parkinson's Disease;
memory loss and/or confusion; psychosis; schizophrenia, schizophrenia with AD
a comorbit with AD], late-onset schizophrenia, paraphreny, schizophrenia-like disorder
anxiety, bipolar disorder, mania; mood stabilization; cognitive impairment after removal of certain gliomas; synucleinopathies (Parkinson's disease, dementia with Lewy bodies, multiple system atrophy); tauopathy; chronic traumatic encephalopathy; Pick's disease; progressive supranuclear palsy; corticobasal degeneration), tardive dyskinesia; sepsis-related encephalopathy; sepsis-induced delirium; memory impairment in disease or schizophrenia; chemotherapy-induced cognitive deficits; alcoholic dementia, post-bypass and transplant cognitive deficits, hypothyroidism-related dementia, autism-related cognitive impairment, Down syndrome, nicotine, Cognitive impairment due to substance abuse or withdrawal, including cannabis, amphetamines, cocaine, attention deficit hyperactivity disorder (ADHD), pain, rheumatism, arthritis, and terminal illness; xerophtalmia, vaginal dryness, dry skin; immunity Dysregulation of food intake, including neuroclinic disorders and hyperphagia and anorexia; obesity; congenital ornithine transcarbamylase deficiency; olivopontocerebral atrophy
]; alcohol withdrawal; substance abuse, including withdrawal and replacement therapy, Huntington's disease; progressive supranuclear palsy; Pick's disease; Friedreich's ataxia; inflammatory bowel disease, irritable bowel syndrome, diarrhea, constipation,
Autonomic disorders, including gastric acid secretion and dysfunction of gastrointestinal motility and function such as ulcers; urge incontinence, asthma, COPD; brain, nervous system, cardiovascular system, immune system, neuroclinic system, gastrointestinal system,
or a dysfunction of one or more of the endocrine and exocrine glands, the eye, the cornea, the lung, the prostate, or other organs whose cholinergic function is mediated by muscarinic receptor subtypes, wherein a brain amyloid-mediated disorder glycogen synthase kinase (GSK3beta)-mediated disorders; tau protein hyperphosphorylation-mediated injury, dysfunction, or disease; CNS and P
NS hypercholesterolemia and/or hyperlipidemia mediated injury, dysfunction or disease; Wnt mediated signaling abnormalities; neuroreversible disorders; hyperglycemia; diabetes; endogenous growth factor mediated diseases; or a central or peripheral nervous system disease state due to impairment involving a combination of additional risk factors; or a disease state involving apolipoprotein E; Delayed onset of AD symptoms in certain patients, Lewy body dementia, Lewy body disease, cerebral amyloid angiopathy (CAA)
, cerebral amyloidosis, frontotemporal dementia, vascular dementia, hyperlipidemia, hypercholesterolemia, multiinfarct dementia (MID), stroke ischemia, MI with stroke/ischemia/head injury
D, mixed MID and Alzheimer's disease, human head trauma, age-related memory impairment, mild cognitive impairment (MCI), MCI leading to AD, bipolar disorder, mania, schizophrenia, nonaffective schizophrenia sychozophrenia], paraphreny, immune dysfunction, neuroclinic disorders, and dysregulation of food intake, including hyperphagia and anorexia, weight control, obesity, and cholinergic dysfunction, including inflammation. Disorders are mentioned, and special interest is aroused in advocating immunotherapy for inflammatory disorders.

[39] Preferably, cholinesterase inhibitors, nicotinic agonists, cholinergic precursors and cholinergic enhancers, nootropics, peripheral antimuscarinic agents [peri
pheral antimuscarinc drug], M2 muscarinic antagonists, M4 antagonists, benzodiazepine inverse agonists, sigma-1 agonists, antidepressants, tricyclic antidepressants or antimuscarinic drugs used in the treatment of Parkinson's disease (PD) or depression agonists, antipsychotics and antischizophrenias, glutamate antagonists and modifiers,
Metabotropic glutamate receptor agonists, NMDA antagonists, AMPA agonists, acetyl-L-carnitine, MAO-B inhibitors, peptides and growth factors, cholesterol-lowering agents, antioxidants, GSK-3 beta inhibitors, Wnt-ligands , PKC activators, beta- or gamma-secretase inhibitors, beta-amyloid degrading agents, neprilysin, insulin degrading enzymes, or activators of endothelin converting enzymes involved in the degradation of beta-amyloid. activators, beta amyloid antiaggregants, chelating agents, antibodies and immunotherapeutic compounds against beta amyloid, antibodies and immunotherapeutic compounds against tau protein pathologies, antibodies and immunotherapeutic compounds against alpha-synuclein pathologies, compounds that bind amyloid, Cyclooxygenase (C
OX)-2 inhibitors, non-steroidal anti-inflammatory agents, estrogenic agents, estrogen receptor modulators, steroidal neuroprotective agents, and spin trap pharmaceuticals. The pharmaceutical composition of [33] or [34], further comprising an additional pharmacologically active compound.

According to the present invention, a pharmaceutical composition based on a crystalline polymorph of Compound A that can be used for treating or preventing the above-mentioned diseases and disorders responsive to stimulation of M1 muscarinic receptors The products can be formulated into various preparations and administered using various routes of administration. Guidance on suitable formulations for various types of administration can be found, for example, in R.
Emington: The Science and Practice of Pha
rmacy (2000) by the University of Science
s in Philadelphia, ISBN 0-683-306472. Accordingly, the present invention also provides methods of administering drugs for treating or preventing diseases or conditions responsive to agonistic stimulation of M1 muscarinic receptors. Accordingly, in further embodiments, the invention relates to:
[40] A method for treating a subject, ie, a patient suffering from a medical condition defined in [37] or [38], comprising a therapeutically effective amount of A method comprising administering the crystalline polymorphic form of any one or the pharmaceutical composition of [33], [34], [38], or [39] to a subject in need thereof. .

The crystalline polymorphic forms of the present invention as characterized above and various aspects and embodiments thereof shall now be described with reference to the following figures and examples. Furthermore, the disclosure content of the above description in the background of the invention forms part of the disclosure of the present invention to the extent applicable. It should be understood that this invention is not limited to the particular methodology, protocols, cell lines, excipients, carriers, and reagents per se described herein. Also, it is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the invention. The scope of the invention shall be limited only by the appended claims.

The following examples help illustrate embodiments of the invention. It will be understood that the invention is not intended to be limited by the foregoing description, which is intended to help illustrate embodiments of the invention.

Methodology Materials Compound A (m.p. 134°C; purity 99.9% by HPLC (achiral); HPLC
(chiral) purity 99.7-100%) were used for crystallization studies.
Water was deionized and passed through an ion exchange system.
All solvents and reagents were analytical grade.
Melting points were recorded on an Electrothermal 9100 capillary melting point apparatus. No correction has been made.

X-ray powder diffraction (XRPD) measurement method and conditions Apparatus: Philips PW-1050/70 type X-ray diffractometer Graphite monochromator Voltage: 40 kv
Current: 28mA
Slit: Receiving slit 0.2 mm, Scattering slit 1 mm
Minimum 2-seater: 3.00
Max 2seater: 40.00
Step size: 0.05
Count time: 0.50 seconds Radiation source: CuK _α
Unless otherwise specified, 2-theta values are listed rounded to 0.1, ±0.2.
For API crystal forms, XRPD was also performed according to USP <941>.

Those skilled in the art will appreciate that XRPD patterns of the same sample (obtained on the same or different instruments) may exhibit variations in peak intensities at different 2-theta values. Those skilled in the art will appreciate that XRPD patterns of different samples of the same polymorph (obtained on the same or different instruments) may also exhibit variations in peak intensities at different 2-theta values. XRPD patterns can be substantially the same pattern even if the peak intensities of the corresponding 2-theta signals are different.

Thermal analysis Measurement methods and conditions for differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) Apparatus: METTLER TG50
Software: METTLER TOLEDO STAR® System Range: 30-200°C
Heating rate: 5°C/min Pan: Aluminum standard 40 μl
Purge gas: Nitrogen with a flow rate of 80 ml/min.
fferential scanning calorimetry] (DSC Q, TA Instruments) and TGA (TGA Q500, TA Instruments), and data analysis was performed with a thermal analyzer (Universal Analysis 2000, TA Instruments).
instruments). For DSC, a heating rate of 3°C/min and a modulated cooling rate of 1°C/min was used over the temperature range of 35-200°C, and for TGA, a heating rate of 10°C/min. A speed of up to 110° C. was used and held for 250-360 minutes. TGA analysis of API samples was performed at a heating rate of 10°C/min to a final temperature of 175°C and analyzed at 115°C.

ZnSe ATR-FT-IR absorption spectrum measurement method and conditions Spectrometer: Nicolet 380
Detector: DTGS KBr
Smart accessories: Smart Multi-Bounce ZnSe HATR
Sample scanning times: 36 times Resolution: 4.000
Unless otherwise specified, IR absorption peaks are listed in units of cm ⁻¹ ±5 cm ⁻¹ .

Solid-state CP/MAS ¹³ C NMR measurement method and conditions Spectrometer: Bruker Advance-500
Spectra were measured with a 4 mm CP-MAS probehead at 125.76 MHz.
All spectra were referenced to tetramethylsilane (TMS) using the carbonyl carbon of glycine (176.03) as a secondary reference.
The rotor frequency was 5.0 kHz.
Two-pulse phase modulation (TPPM) was used for proton decoupling.
The contact time was 1000 μs.
4k data points were acquired in 40ms.
The recycle delay was 5.0 seconds for all experiments.
Unless otherwise specified, NMR peaks are rounded to 0.1 ± 0.1, T
Chemical shifts are listed in ppm relative to MS.

Single Crystal X-Ray A single crystal of the compound to be tested was attached to a glass fiber with epoxy glue and transferred to a Bruker SMART APEX CCD X-ray diffractometer equipped with a graphite monochromator. Pentium series P running SMART software package ¹
C controlled the system. The data are presented using MoK _α radiation (λ=0.71073 Å),
Collected at 173K. Immediately after collection, raw data frames were transferred to a second PC computer for integration and reduction with SAINT program package ² . The structure is SHEL
Solved and refined with XTL software package ³ .
1. SMART-NT V5.6, BRUKER AXS GMBH, D-76181
Karlsruhe, Germany, 2002
2. SAINT-NT V5.0, BRUKER AXS GMBH, D-76181
Karlsruhe, Germany, 2002
3. SHELXTL-NT V6.1, BRUKER AXS GMBH, D-761
81 Karlsruhe, Germany, 2002

Microscopy of various crystals was performed using a Nikon TMS inverted optical microscope. Micrographs were obtained using a digital camera (Nikon Japan, MDC lens). Crystals were imaged on glass slides.

Scanning electron microscopy (SEM) of various crystals from ×25 to ×300 at 20 kV
Evaluation was made in the magnification range up to 0.

For hygroscopic APIs, water uptake was measured using a dynamic water uptake experiment (SGA-100 Symmetri
c Vapor Solution Analyzer, VTI Corporation
n). The dynamic water uptake profile of API (Form III) ranges from 40% to 9
Adsorption was completed in 10% increments to 0% RH, followed by 90% to 10% in 10% increments, 10% to 2% in 1% increments, and desorption to 2%. The dynamic water uptake profile of API (Form II) completed adsorption in 10% increments from 40% to 90% RH, followed by desorption to 10%.

Moisture API moisture was obtained from a Settler Toledo DL35 Karl Fische
determined by r Titrator.

Water Solubility Water solubility was performed by preparing 50 mg of API monohydrate (Form III) in 2 ml of deionized water. Samples were prepared in triplicate and the preparations were rotated at room temperature for 2 days. After spinning, the mixture was allowed to stand for 3 hours to allow any undissolved API to settle. The supernatant was filtered using a 4 mm 0.45 microM PTFE filter.
Filtered into a scintillation vial. Pipette 25 microliters and add 97
A 1 in 40 dilution was made by transferring to 5 microliters of sample solvent. The resulting dilutions were placed in vials for HPLC analysis.

Example 1 Crystalline Form I Compound A
a) Ethyl acetate (7 ml) was added to compound A (1.37 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to
1.06 g of solid compound A was obtained. X-ray powder diffraction yielded the following 2-theta values, d-spacings, and relative intensities, confirming that the material was crystalline (Fig. 1).

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 13.269, 30.147, 4.
3.613, 40.968, 52.151, 54.081, 46.585, 67.088
, 174.360.

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 673, 724, 774, 808, 826, 945, 984, 1012
, 1026, 1070, 1110, 1144, 1194, 1278, 1291, 1352
, 1370, 1388, 1426, 1438, 1467, 1676, 2685, 2791
, 2802, 2842, 2921, 2941, 2974, 3023, 3159, 3429
. (Fig. 10)

The DSC of Compound A Crystalline Form I (Figure 3A) corresponds to an endothermic peak at 107.1°C (starting at 104.8°C), corresponding to the release of water that had been physically occluded in the crystal structure, and a melting point. It exhibits an endothermic peak at 134.6°C (starting at 133.8°C). Thermal decomposition occurred after melting. In this case, the broad low temperature peak at 61-77° C. characteristic of Form III (see FIG. 4)
was not observed. Further information on crystalline Form I is provided in Example 27. Form I TGA exhibited a % weight loss of 5.2% when heated to temperatures up to about 115°C. This indicates less than the stoichiometric percent of water required for the complete monohydrate of Compound A (for the stoichiometric monohydrate, the theoretical number of water is 7.76 %).

b) Form I can be obtained by slow evaporation of compound A dissolved in ethyl acetate at 25°C. XRPD showed that the compound was crystalline and had the XRPD pattern of Form I as shown in FIG.
. 9° C.) and 135.8° C. (onset 133.9° C.) exhibiting only characteristic endotherms of Form I.

Example 2 Crystalline Form I Compound A
a) Water (25 ml) was added to compound A (1.07 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature and the precipitated crystals were collected by filtration. The obtained crystals were dried to obtain 0.82 g of Compound A. m. p. is 134.7 to 135.2°C
.

X-ray powder diffraction (XRPD): [2Th, d(A), I/I0] (8.8, 10.052
, 6), (9.6, 9.174, 4), (12.3, 7.223, 73.9), (15.
6, 5.696, 4.9), (17.3, 5.126, 81.3), (17.5, 5.0
66, 100), (19.3, 4.6, 1.8), (19.9, 4.461, 20.3)
, (21.6, 4.106, 39.8), (23.1, 3.857, 5.6), (23.
5, 3.782, 6.3), (24.5, 3.632, 12.2), (26.3, 3.3)
88, 20.9), (27.2, 3.276, 0.5), (28.8, 3.097, 8.
4), (30.3, 2.945, 3.6), (31.2, 2.865, 9.8), (31
. 6, 2.83, 14.1), (32.5, 2.751, 2.6), (34, 2.634
, 3.3), (34.5, 2.597, 5.9), (35, 2.564, 4.8), (3
5.4, 2.533, 9.3), (36, 2.493, 5.9), (37.4, 2.40
5, 2.4), (38.5, 2.338, 4.1), (39.4, 2.285, 3.8)
.

Solid state CP/MAS ¹³ C NMR chemical shift (δ _c in ppm) 13.289, 3
0.148, 43.636, 41.037, 52.164, 54.139, 46.605
, 67.082, 174.406.

ATR-FT-IR absorption peaks (cm ⁻¹ ): 673, 724, 774, 808, 82
6, 890, 944, 984, 1012, 1026, 1070, 1110, 1144, 1
193, 1278, 1290, 1352, 1369, 1388, 1426, 1438, 1
467, 1681, 2685, 2790, 2841, 2888, 2920, 2940, 2
974, 3021, 3159, 3424.

DSC: 105.6°C (onset 104.4°C), endotherm at 134.7°C (onset 133.4°C), very broad trace at 75-95°C.

This polymorph of TGA exhibited a weight loss of 5.2% when heated to temperatures up to about 120°C. This indicates less than the stoichiometric percent of water required for the complete monohydrate of Compound A. This form is defined as Form I with trace amounts of Form III due to characteristic peaks in DSC, XRPD, CP/MAS ¹³ C NMR, and ATR-FT-IR.

b) Form I can be obtained by slow evaporation of compound A dissolved in water at 50°C. XRPD shows the compound to be crystalline, DSC is 107.4
C. (onset 104.8.degree. C.) and 133.9.degree.

Example 3 Crystalline Form I Compound A
Isopropanol (3.5 ml) was added to compound A (1.0 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to give 0.5 g of compound A as a solid.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 13.423, 30.301, 4.
3.729, 41.122, 52.289, 54.237, 46.739, 67.222
, 174.457.

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 725, 775, 809, 827, 890, 945, 985, 1012
, 1027, 1071, 1111, 1144, 1195, 1278, 1292, 1353
, 1371, 1389, 1427, 1438, 1468, 1672, 2845, 3021
, 3158, 3427.

This form is designated Form I due to the characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 4 Crystalline Form I Compound A
Tetrahydrofuran (THF, 0.5 ml) was added to compound A (0.18 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to yield compound A as a solid.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 13.355, 30.227, 4.
3.679, 41.090, 52.225, 54.170, 46.678, 67.153
, 174.525.

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 674, 726, 775, 809, 827, 985, 1027, 107
1, 1111, 1144, 1195, 1278, 1292, 1353, 1371, 138
8, 1426, 1438, 1468, 1676, 2791, 2850, 2920, 294
0, 2974, 3023, 3159, 3430.

This form is designated Form I due to the characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 5 Crystalline Form I Compound A
Ethanol (0.5 ml) was added to Compound A (0.11 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to
A solid compound A was obtained.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 13.304, 30.191, 4
3.674, 40.999, 52.180, 54.138, 46.637, 67.100
, 174.426.

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 673, 724, 774, 808, 826, 985, 1027, 104
6, 1070, 1110, 1278, 1291, 1353, 1370, 1388, 142
6, 1438, 1467, 1669, 2790, 2842, 2887, 2920, 294
0, 2974, 3022, 3161, 3426.

This form is designated Form I due to the characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 6 Crystalline Form I Compound A
tert-butyl methyl ether (1.3 ml) was added to compound A (0.11 g),
The mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to yield compound A as a solid.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 13.134, 30.049, 4.
3.479, 40.789, 53.979, 51.999, 46.448, 66.927
, 174.217.

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 725, 775, 809, 827, 985, 1027, 1071, 11
11, 1144, 1195, 1278, 1292, 1353, 1371, 1388, 14
27, 1438, 1468, 1670, 2790, 2842, 2974, 3021, 31
57, 3430.

This form is designated Form I due to the characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 7 Crystalline Form II Compound A; API
a) Pure Compound A crystalline Form II can be formed by recrystallization from acetone (Figures 6-8). XRPD of Form II is shown in FIG.
It differs from the XRPD of II (FIGS. 1 and 2). DSC showed only one endothermic peak at 135.35° C. (onset 134.2° C.; FIG. 7A).

b) Pure crystalline Form II was also prepared as kg quantities of the cGMP compound and used as an API in preclinical and clinical studies. As shown by XRPD in FIG. 6B and Table 1, this API is crystalline. DSC of this API showed an endothermic peak at 134.29° C. (FIG. 7B). TGA of anhydrous crystalline Form II of Compound A showed no significant weight loss up to 110° C. (FIG. 8). Further information on Form II can be found in Example 25
and 26.

c) Pure anhydrous crystalline Form II can be obtained from a saturated solution of Compound A prepared by adding Compound A to acetone at 30°C to 50°C. These were mixed with acetone/
Precipitation was induced by rapid cooling in an ice bath. The solid that formed was isolated and characterized by XRPD. XRPD showed characteristic peaks of Form II.

d) Pure anhydrous crystalline Form II can be obtained from saturated acetone solutions of Compound A prepared at 30° C. and 50° C. slowly cooled in a programmed circulating bath.
The slurry formed was then heated to 50° C. for 2 hours and then cooled to 25° C. for 2 hours. This process was repeated overnight and a solid was isolated for further analysis by XRPD. XRPD showed characteristic peaks of Form II.

Example 8 Crystalline Form II Compound A
a) To compound A (1.05 g) was added acetonitrile (9 ml) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to give 0.71 g of compound A as a solid. X-ray powder diffraction yielded the following 2-theta values, d-spacings, and relative intensities, confirming that the material was crystalline.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 15.371, 14.834, 2
8.871, 26.010, 52.326, 49.327, 43.582, 39.487
, 38.761, 45.773, 44.974, 65.289, 64.013, 175.
005.

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 729, 773, 822, 906, 944, 989, 1027, 106
5, 1108, 1147, 1194, 1279, 1294, 1340, 1362, 142
5, 1447, 1464, 1667, 2792, 2850, 2869, 2901, 294
0, 2951, 3006, 3160, 3428.

DSC is 50-77°C (very small and broad, probably due to the presence of traces of Form III), 105.2°C (starting at 104.5°C; very low, probably due to the presence of traces of Form I), and It exhibited an endotherm at 134.8°C (onset 133.4°C).

b) Pure anhydrous crystalline Form II can be obtained with dry acetonitrile (Example 2
6 and Figure 11).

c) Pure anhydrous crystalline Form II can be obtained from slurries of monohydrate Forms I and III in acetonitrile (see Example 22).

d) Pure anhydrous crystalline Form II can be obtained from a saturated solution of Compound A prepared by adding Compound A to acetonitrile at 30-50°C. This was rapidly cooled in an acetone/ice bath to induce precipitation. The solid that formed was isolated and characterized by XRPD showing the characteristic peaks of Form II.

e) Pure anhydrous crystalline Form II can be obtained from saturated acetonitrile solutions of Compound A prepared at 30° C. and 50° C. slowly cooled in a programmed circulating bath. The slurry formed was then heated to 50° C. for 2 hours and then cooled to 25° C. for 2 hours. This process was repeated overnight and a solid was isolated for further analysis by XRPD. XRPD showed characteristic peaks of Form II.

Example 9 Crystalline Form II Compound A
a) Hexane (100 ml) was added to compound A (1.25 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to give 1.06 g of compound A as a solid. X-ray powder diffraction confirmed that this material was crystalline Form II with traces of Form I. This form is CP/MAS ¹³ CN
It is defined as Form II due to the characteristic peaks in MR and ATR-FT-IR.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave a spectrum with the following chemical shifts (δ _c in ppm, fractions kept): 15.385, 14.829, 2
8.998, 25.945, 52.293, 49.345, 43.595, 39.504
, 45.791, 45.014, 65.441, 64.086, 174.963 (see Table 6).

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions preserved): 730, 773, 805, 820, 854, 905, 944 ^* , 988
^* , 1027 ^* , 1064, 1108 ^* , 1148, 1193, 1278, 1294, 1
339, 1362, 1424, 1446, 1458, 1664, 2685, 2723, 2
771, 2850, 2868, 2901, 2940, 2951, 3005 ( ^* These peaks are due to minor amounts of Form I in Compound A Form II).

b) Pure anhydrous crystalline Form II can be obtained by phase equilibrating a hexane suspension of Compound A at 50° C.±1. After equilibrating for 24 hours, the supernatant is filtered,
Solids were collected and analyzed by XRPD. X-ray powder diffraction confirmed that the material was crystalline, yielding characteristic peaks of Form II 2-theta values, d-spacings, and relative intensities.

Example 10 Crystalline Form II Compound A
Dioxane (4 ml) was added to compound A (1.48 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to give a 0.
77 g of solid compound A were obtained. X-ray powder diffraction yielded the following 2-theta values, d-spacings, and relative intensities, confirming that the material was crystalline.

Solid phase CP/MAS ¹³ CN of crystalline material as shown in FIG. 9 (Form II)
MR gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 15.359, 14.788, 28.926, 25.956, 52.311, 4
9.342, 43.462, 39.543, 45.806, 44.960, 65.557
, 63.967, 174.965.

As shown in FIG. 10 (Form II), ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 729, 773, 822,
906, 944, 988, 1027, 1065, 1109, 1148, 1194, 127
9, 1294, 1339, 1362, 1426, 1447, 1471, 1671, 279
3, 2850, 2869, 2901, 2941, 2951, 3012, 3159, 343
2.

DSC is 56-66° C. (very small, possibly residual solvent or traces of Form III)
and 134.9°C (starting at 133.7°C).

This form is also designated as Form II due to characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 11 Crystalline Form II Compound A
Diethyl ether (7 ml) was added to compound A (0.19 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to yield compound A as a solid.

Solid state CP/MAS ¹³ C NMR of the crystalline material gave a spectrum with the following chemical shifts (δ _c in ppm, fractions kept): 15.206, 14.629, 1
3.102 ^* , 28.814, 25.747, 52.167, 49.182, 43.32
6, 39.323, 38.529, 45.610, 44.756, 66.827 ^* , 65
. 310, 63.845, 174.863. ( ^* Peaks at 13.102 and 66.827 may be due to the presence of trace amounts of Form I).

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 728, 773, 822, 944, 987, 1027, 1065, 11
08, 1147, 1278, 1293, 1339, 1362, 1425, 1445, 14
59, 1664, 2787, 2847, 2869, 2902, 2921, 2951, 30
06, 3147.

This form is designated as Form II due to the characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 12 Crystalline Form II Compound A
Pure anhydrous crystalline Form II was slowly cooled in a programmed circulating bath, 3
It can be obtained from saturated hexane solutions of compound A prepared at 0°C and 50°C. The slurry formed was then heated to 50° C. for 2 hours and then cooled to 25° C. for 2 hours. This process was repeated overnight and the solid was isolated for further analysis by XRPD and showed characteristic peaks of Form II.

Example 13 Crystalline Form II Compound A
Cyclohexane (5 ml) was added to compound A (0.10 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to
A solid compound A was obtained.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 15.356, 14.775, 2
9.061, 25.940, 52.340, 49.360, 43.368, 39.704
38.721, 45.819, 44.950, 65.646, 64.006, 175.0
93.

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 668, 682, 707, 749, 774, 809, 824, 862,
945, 988, 1027, 1065, 1110, 1146, 1195, 1279, 12
94, 1351, 1448, 1468, 1676, 2735, 2792, 2816, 28
27, 2851, 2900, 2922, 2941.

This form is designated as Form II due to the characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 14 Crystalline Polymorphic Form III Compound A
Crystalline Form III Compound A can be produced from reslurrying Compound A in deionized water and filtering. X-ray powder diffraction confirmed that the material was crystalline (see Figure 2).

Initial experiments attempted to identify the solvent used to recrystallize Compound A for large-scale cGMP crystallization of API show that a variety of solvents were used to produce API in good recovery. showed that it is possible. However, many of these solvents (especially ethyl acetate/heptane, ethanol, tetrahydrofuran, toluene, isobutyl acetate, methyl isobutyl ketone), due to the fact that long needles of the final product stuck to the sides of the flask, caused large scale was not suitable for conversion. Manual scraping was required to remove these sticky solids. Acetone was found to be one solvent that produced a white material with minimal loss of solids to the sides of the flask. In this case, recrystallization from acetone gives
Crystal Form II was produced (Figures 6-8). The final requirement required to produce Form III was hydration of the final product. Experimental work has shown that compound A can absorb about 1 molar equivalent of water in aqueous acetone when stirred for approximately 0.5 hours. Therefore, by adding 1.3 molar deionized water to an acetone solution of Compound A, large kg quantities of crystalline Form III Compound A were produced as a cGMP API. Since Compound A (Form II) can readily absorb water, it is necessary to develop a drying method that will remove acetone without removing water. Oven drying experiments proved to be an impractical way of doing this. This material was subjected to a full house vacuum at 40°C.
Drying under the hood overnight removes all water as shown by Karl Fischer analysis. Even at 20-25° C., water could be removed from the molecule under full house vacuum. It was later discovered that water was removed when the relative humidity dropped below 10%. Conversely, when the relative humidity exceeded 60%, the product regained its water and eventually became fully hydrated. From this data, two hydration methods were developed. Both methods first form a hydrate by adding 1.3 moles of water to a homogeneous crystallization solution; filtering the hydrated material; analyzing by Karl Fischer; This includes drying the material on the filter with vacuum and nitrogen flow. This removes the acetone without dehydrating the molecule. The other method may have included using an ambient temperature vacuum oven to remove the acetone and then rehydrating the material by increasing the relative humidity within the oven to 60-90%. 15.1 l of acetone, 1 kg
of Compound A was charged to the vessel and the solution was subjected to atmospheric reflux. The solution is then held at reflux to ensure complete dissolution of the solids. The solution is then cooled to 35-45° C. and passed through an in-line filter to remove any particulates. The solution is now heated and distilled under atmospheric pressure to 7.4-7.6 l. 0.1 l of deionized water is added and the solution is cooled to -5 to -10°C over 3-4 hours. The solution is then held at -5 to -10°C for 1 hour or more. An in-process control is taken at this point and the filtered solids are analyzed for moisture content and if the Karl Fischer analysis is higher than 7.5%, proceed with the process. The solids are then filtered and dried on the filter under vacuum and _N2 flow for a minimum of 4 hours. Crystallization yields approximately 1.0 kg of Compound A monohydrate, Form III, as API (~93
% yield) (see Figure 4B, Figure 5, and Example 24).

X-ray powder diffraction confirmed that the material thus obtained was crystalline (
See Table 2 and Figure 2).

Figure 4. Differential scanning calorimetry (DSC) of the monohydrate crystalline form of Compound A (Form III).
represents a curve. A) From a reslurry in water. DSC shows two endothermic peaks, one at about 77.10°C and the other at about 134.87°C. B) cGMP active pharmaceutical ingredient (API) prepared by crystallization from acetone and 1.3 equivalents of water. The DSC is about 61.1 on the one hand.
It shows two endothermic peaks at 8°C and another at about 133.75°C.

Figure 5 shows a thermogravimetric analysis (TGA) of the monohydrate crystalline form of Compound A (Form III, API).
). When heated to temperatures up to about 110°C, the TGA shows a weight loss of 7.8%. Moisture by Karl Fischer analysis is 7.7% as expected from a stoichiometric monohydrate.

Example 15 Conversion of Form II to Form III Compound A crystalline Form II (0.4631 g; crystallized from acetonitrile) was placed in a room temperature desiccator ₍ saturated solution of _Na2HPO4 ) at 95% relative humidity for 1 week. (Fig. 1
5A). X-ray powder diffraction, ATR FT-IR, CP-MAS solid-state ¹³ C NMR, DSC
, and TGA, showed that the material was Form III, not crystalline Form I Compound A or Form II Compound A (see Figure 15B). Form III
It will be seen that the X-ray powder diffraction, ATR FT-IR, and ¹³ C NMR characteristics of Form I are similar to those of Form I. However, DSC examination showed endotherms at 58-94° C. (very broad) and 133.9° C. (onset 133.4° C.) and the TGA curve was
It showed a 7.3% loss in weight from 58°C to 94°C, indicating the presence of one molecule of water for every molecule of Compound A. Both of these are features of Form III (FIG. 15). Remarkably, such continuous dehydration behavior with relatively low temperature onset is also common to channel hydrates to which Form III is assigned (Mirza et al., AAPS Pharma S
ci. 5(2):2003). X-ray powder diffraction confirmed that the material thus obtained was crystalline.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave spectra with the following chemical shifts (δ _c in ppm, fractions kept): 13.771, 30.702, 4.
4.121, 41.489, 54.598, 52.647, 47.073, 67.559
, 174.932.

ATR-FT-IR gave a spectrum with the following absorption peaks (cm ⁻¹ , fractions kept): 725, 774, 808, 827, 985, 1039, 1070, 11
10, 1144, 1194, 1277, 1291, 1353, 1369, 1388, 14
26, 1438, 1467, 1659, 2789, 2835, 2918, 3088, 31
44, 3422.

Example 16 Mixture of Forms I and II To compound A (0.12 g) was added dichloromethane (0.2 ml) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to yield compound A as a solid.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave a spectrum with the following chemical shifts (δ _c in ppm, fractions kept): 14.708, 14.581, 1
3.122, 30.010, 25.692, 53.942, 52.012, 49.300
, 43.437, 40.832, 46.449, 66.949, 65.522, 63.8
84, 174.237.

This form is defined as a mixture of Form I and Form II due to the characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 17 Mixture of Forms I and II Toluene (0.5 ml) was added to compound A (0.12 g) and the mixture was heated to dissolve the compound. The clear solution was cooled overnight at room temperature. A solid material formed in the solution that appeared to be crystalline to the naked eye. This material was collected by filtration and dried under vacuum at room temperature to yield compound A as a solid.

Solid-state CP/MAS ¹³ C NMR of the crystalline material gave a spectrum with the following chemical shifts (δ _c in ppm, fractions kept): 15.331, 14.724, 1
3.275, 30.157, 29.055, 25.906, 54.082, 52.728
, 52.228, 49.334, 43.489, 39.538, 37.700, 45.7
90, 44.884, 67.063, 65.623, 63.970, 175.004.

This form is defined as a mixture of Form I and Form II due to the characteristic peaks in CP/MAS ¹³ C NMR and ATR-FT-IR.

Example 18 Conversion of Form I to Form II by Heating a) Compound A crystalline Form I (crystallized from ethyl acetate) was heated at 90° C. while Form I
Although the characteristic peaks of (region 14 of the solid-state CP/MAS ¹³ C NMR spectrum
-15, 26-29, 44-46, and 63-66 ppm), and the characteristic peaks of Compound A Crystalline Form II increased [distinguishing peaks (15.4, 14.7), (29.1
, 25.9), (64.0, 65.7) ppm]. Compound A crystalline Form I was completely converted to Compound A crystalline Form II in 4 hours.

b) Crystalline Form I/III Compound A crystallized from water was heated at 90° C. for 75 minutes. Solid-state CP/MAS ¹³ C NMR of the product confirmed the conversion of crystalline Form I/III to crystalline Form II (see Figure 16).

c) Crystalline Form II Compound A was heated at 70° C. for 10 hours and then left at room temperature overnight. Solid-state CP/MAS ¹³ C NMR of the product confirmed that crystalline Form II was unchanged.

Example 19 Conversion of Form I to Form II by Melting a) Heat crystalline Form I/III Compound A (crystallized from water) to 160° C., then melt the material at room temperature for crystallization. I left it as is. Product Solid Phase CP/MAS ^1
3 C NMR confirmed that crystalline Form II was obtained.

b) Crystalline Form II Compound A (crystallized from acetonitrile) was heated to 160° C. and then the molten material was left at room temperature for crystallization. Solid phase CP/MA of the product
S ¹³ C NMR confirmed that crystalline Form II was obtained.

Example 20 Conversion of Form II to Form I or to a Mixture of Forms I and III, respectively under ambient conditions a) Crystalline Form II Compound A was allowed to stand at room temperature for 90 days. Crystalline Form I
The conversion of I to crystalline Form I was confirmed by solid-state CP-MAS ¹³ C NMR.

b) API Form II converts to Form I after exposure to 90% relative humidity for 3 hours as evidenced by Modular DSC (MDSC). Therefore, Form I
Form I (MDSC=104.0°C and 133.8°C)
).

c) API Form II was obtained at MDSC (isothermal lines: 62.2°C, 104.2°C, and 134°C).
. 1° C.), it transforms into a mixture of Forms I and III upon exposure to ambient humidity for 4 months at room temperature.

Example 21 Crystalline Form I Does Not Convert to Form III at _{High Humidity} saturated solution) for 1 week. Analysis of the crystalline material by X-ray powder diffraction, ATR FT-IR, CP-MAS solid-state ¹³ C NMR, DSC, and TGA indicates that the material is still crystalline Form I and Form III
It was confirmed that it was not converted to This is another indication that Form I and Form III are different crystalline forms.

Example 22 Slurry Conversion Acetonitrile (2 ml) was added to a mixture of crystalline Forms I and III of Compound A (0.1
5 g, crystallized from water) and the resulting slurry was stirred at room temperature for 6 hours, after which the residual solids were filtered. Solid-state CP-MAS ¹³ C NMR confirmed the conversion of the mixture of crystalline Forms I and III to crystalline Form II.

Example 23 Phase Equilibration of a Mixture a) A given amount of an anhydrous crystalline form and a monohydrate crystalline form of Compound A is combined with a given amount of either the anhydrous or monohydrate form of Compound A. Individually mixed, then equilibrated in acetone and monitored by XRPD. Both experiments yielded a substantially pure anhydrous crystalline form (Form II).

b) individually mixing predetermined amounts of the anhydrous crystalline form and the monohydrate crystalline form of Compound A with predetermined amounts of either the anhydrous or monohydrate form of Compound A, followed by equilibration in water; let
Monitored by XRPD. The solid is substantially pure monohydrate Form II within 2 hours
completely transformed into I.

Example 24 Physical Properties of Compound A Monohydrate Crystalline Form III Physical properties of Compound A monohydrate crystalline Form III are summarized in Tables 2, 3 and Examples 14 and 21. there is

An X-ray powder diffraction pattern shows the sample to be a crystalline material (Figure 2). D of this sample
SC showed about 77.1° C. (broad) on one side and about 134.9° C. on the other (FIG. 4A), and for API, 61.18° C. (broad) and 133.75° C. (FIG. 4B). Two DSC peaks are shown. TGA results (Figure 5) show that there was a weight loss of 7.8% up to about 110°C, as expected from a stoichiometric monohydrate. Thermal decomposition occurred immediately after melting and the material was completely decomposed around 230°C.

cGMP API Form III is highly stable under long-term conditions of 25° C./60% RH, appearing (white powder) with DSC (T=0, 64, as expected from the features of Form III).
. 2° C. and 133.8° C., and T=12 months, 64.5° C. and 133.8° C.) with 100% purity by HPLC (achiral) and 99.9.
From 9 to 100%, there is no change in water content (7.6-8%). Form III does not convert to Form I. Furthermore, Form I is not converted to Form III. This is another indication that Form I and Form III are different, non-interconvertible crystalline forms.

Importantly, API Form III is physically stable and non-hygroscopic. 8~
No water was absorbed at 90% relative humidity. As a result, the powder pattern also does not change.
No water desorption was detected above 7% relative humidity. Form III begins to dehydrate below 7% relative humidity and virtually complete water loss is achieved at 2% relative humidity.

Aqueous solubility of API (form III) = 5.3 mg/ml.

Example 25 Physical Characteristics of Compound A Anhydrous Crystalline Form (Form II) Physical characteristics of Compound A anhydrous crystalline Form II are summarized in Tables 1 and 4.

The X-ray powder diffraction pattern (Fig. 6, Table 1) indicates that the sample is crystalline, approximately 131.2-13
It is shown to have a melting point of 3.3°C. DSC indicates that the crystalline material has a T _onset of about 134.2°C.
, and a T
It shows that it has a _peak . TGA results (Figure 8) show no significant weight loss below 110°C. After melting, thermal decomposition occurred and the material was completely decomposed at about 250°C.

API Form II is highly stable for over two years as anhydrous Form II if maintained under a dry atmosphere. Anhydrous Form II begins to convert to the hydrate form upon storage at ambient room temperature and humidity. Evidence for partial conversion of the anhydride to the hydrate is based on the increase in water content observed upon repeated handling. When anhydrous Form II is equilibrated at high humidity, conversion to monohydrate Form III occurs based on stoichiometric molar water uptake and thermal properties.

Solid-state CP/MAS ¹³ C NMR can also be used to ascertain the number of molecules in the crystallographic asymmetric unit (Harris, Analyst 131 (2006), 351
-373; Harris, Solid State Sciences 6 (2004)
, 1025-1037). Molecular crystals containing more than one homogenous molecule in an asymmetric unit exist in crystallographically distinct sites and therefore have different environments. Consequently, NMR will show that they have different properties, and similar atoms (eg, carbon) will, in principle, differ in their chemical shifts. Generally, the ^peak is
When appearing as multiplets of C resonances, the number of components of such multiplets indicates the number of molecules in the asymmetric unit. Thus, for Compound A crystalline Form I (see Figure 9B and Table 5), one set of signals was observed. This indicates that one molecule of Compound A resides in the asymmetric unit (see also Example 27, Figure 14). On the other hand, for compound A crystalline form II (
9A and Table 5), doubling of some resonance peaks (especially regions 14-15, 26
29, 44-46, and 63-66 ppm) indicates that more than one molecule of Compound A is present in the crystalline unit (see also Example 25, Figure 11). Solid-state CP/MAS ¹³ C NMR spectra of each form alone (Compound A Crystalline Forms II and I, respectively) [Fig. 9A, distinctive peaks (15.4, 14.7), (29.1, 25.9) , (64.
0, 65.7) ppm; Figure 9B, distinctive peaks 13.3, 30.2, 67.1 ppm] and the spectrum of the true mixture of crystalline forms I and II [Figure 9D, distinctive peaks (15.
4, 14.7, 13.3), (29.1, 25.9, 30.2), (64.0, 65.7)
, 67.1) ppm] clearly shows that Compound A crystalline Form II does not contain detectable amounts of crystalline Form I. Furthermore, Compound A crystalline Form II does not contain detectable amounts of Compound A crystalline Form III (Figures 9C and 9D). Notably, Form I and Form III have the same CP/MAS ¹³ C NMR spectrum (Fig. 9B
and 9C).

Inverted optical microscopy showed that the crystals obtained from ethyl acetate (Form I), water, and ethanol were plate-like crystals, whereas the crystals obtained from isopropanol showed flat crystals.
Crystals obtained from acetone, tetrahydrofuran, and tert-butyl methyl ether show needle-like crystals. Anhydrous Form II of Compound A can be acetonitrile, cyclohexane,
Crystallized as needles from hexane and diethyl ether.

Scanning electron microscopy (SEM) showed anhydrous Form II and monohydrate Form I of Compound A
II are long rod-shaped crystals. However, the Form III monohydrate lot appears to have a more uniform crystal size distribution. Anhydrous samples appear to have a few long crystals in mostly smaller crystals. The surface texture of the anhydrous form appears smoother compared to the monohydrate crystal surface at higher magnification.

Example 26 Single Crystal X-Ray of Compound A Form II (Crystallized from Acetonitrile) A single crystal of Compound A was attached to a glass fiber with epoxy glue and Bruker SMART APEX CCD X-ray diffraction with a graphite monochromator. moved to meter.
Data were collected at 173 K using MoK _α radiation (λ=0.71073 Å) and the SMART software package. Immediately after collection, raw data frames were transferred to the SAINT program package for integration and reduction. Structures were solved and refined with the SHELXTL software package (all software was Bruker AXS
GmbH, Karlsruhe, Germany). Compound A anhydrous crystal single crystal X
Line data are summarized in Tables 6-11.

Compound A anhydrous crystals were crystallized in space group P2(1) with four molecules in the unit cell. There are two Compound A molecules in the asymmetric unit of the P2 ₁ chiral space group anhydride crystal (
CP/MAS ¹³ C NMR studies so conclude). These two molecules are conformationally different. Molecule 1 has a five-membered ring staggered conformation with a pseudo _C2 axis due to the C=O bond. Molecule 2 has a five-membered ring envelope conformation with the S atom occupying the flap position. FIG. 11 shows the molecules and their positions in the unit cell of the structure.

Computer Simulations of Anhydrous Single Crystals of Compound A and Crystalline Material Form II
showed that both were the same polymorph (Fig. 12).

Example 27 Single Crystal X-Ray of Compound A Crystal Form I (Crystalized from Ethyl Acetate) Compound A single crystal Form I has four molecules in the unit cell, space group P2(1)2(1).
2(1) was crystallized. There is one compound A molecule in the asymmetric unit of the monohydrate crystal measured at low temperature [173(1) K]. FIG. 14 shows the molecules and their positions in the unit cell of the structure. The molecule has a five-membered ring, twisted conformation, and a pseudo _C2 axis due to the C═O bond. There are two strong hydrogen bonds in compound A crystal form I (crystallized from ethyl acetate): amide N(1)...O(1w) 2.757 Ang, amide H(1)...O(1
w) 1.947 Ang, and N(1)-H(1)...O(1w) 172.88 degrees; and O(1w)...O(1) carbonyl 2.735 Ang, water H(1w)・・・O(
1w) 1.945 Ang, O(1w)-H(1w)...O(1) 165.38 degrees. There is also one weak hydrogen bond: piperidine N(2)...O(1w)2.852An
g, water H (2w) ... N (2) 2.302Ang, O (1w) - H (2w) ... N (
2) 118.36 degrees; Without wishing to be bound by theory, the inventors believe that this hydrogen-bonded water molecule is the one responsible for the DSC/TGA peak at 104°C.

FIG. 13 is a single crystal form of XPRE shown in US Pat. No. 7,439,251 B2.
3 represents three X-ray powder diffraction patterns of Compound A Forms I, II, and III, respectively, simulated by using P. Notably, compound A crystalline form I
and crystalline form III are similar, although U.S. Pat. No. 7,43
There are still some differences in the diffraction angles (see Table 13, Diffraction angles of peaks marked with ^* are >+
/-2 degrees). Therefore, XRPD of Form I and Form III can be obtained from US Pat.
, No. 251 B2 of the single crystal of compound A hydrate published in XPREP,
This is further confirmation that Form I and Form III are distinct, unpublished novel polymorphic forms (Table 13). Furthermore, the XRPD of polymorphic Form II shows that X of a single crystal of this form
Identical to the RPD simulation (FIG. 12), but US Pat. No. 7,439,251B
2 and from XRPD of both Form I or Form III (FIG. 13).

Example 28 Form II and Form II in vials for reconstitution as an oral solution
Formulation of Each Compound A Form II and Form III powders were each added to 8 ml of amber glass (Form I).
Fill vials (120 mg/vial) and close the vials with Teflon/rubber screw caps. Addition of approximately 5 ml of distilled water and complete dissolution of the crystalline powder results in an oral solution that can be used, inter alia, to treat the diseases mentioned above.

Example 29 Formulation of Form III in Oral Capsules Compound A Form III powder is mixed with one or more excipients (pregelatinized starch, microcrystalline cellulose, colloidal silicon dioxide, and stearic acid). and the mixture was added to size 4 to provide 5 mg or 10 mg of Compound A Form III per capsule.
white opaque hard gelatin bisected capsules. This capsule can be used as an oral formulation for immediate release in the gastrointestinal tract.

Example 30 Formulation of Form II in Oral Tablets Compound A Form II powder was mixed with one or more anhydrous excipients (e.g., anhydrous calcium hydrogen phosphate), and 5 mg, 10 mg per tablet, Or directly compress into tablets containing 20 mg of active agent.

Example 31 Long-Term and Accelerated Bulk Stability of API, Compound A Monohydrate (Form III) Highly fixed API: Compound A (NGX267) clinical batch (cGMP) at 25°C
/60% RH meets long-term specification at 0, 3, 6, 9, 12, 18, and 24 months, appearance (white powder), DSC (T=0, 64 .2, and 133
. 8° C.; and T=12 months, 64.5 and 133.8° C.), purity by HPLC (achiral)=100% and purity by HPLC (chiral) (99.9-100%), and water content (7.2-8%) did not change.

Highly fixed API in formulation: Capsule drug product containing 5 mg of Compound A (NGX267) and Capsule drug product containing 10 mg of Compound A (NGX267) at 25°C/60°C.
It met specifications for up to 18 months when stored at % RH and up to 6 months when stored at 40°C/75% RH.

Example 32 Pharmacokinetics of Form II in Humans In a first Phase I clinical study, an orally administered API was prepared according to Example 29 but with additional higher and lower dosages. A total of 34 subjects were randomized to receive study drug (1, 2.5, 5, 10, 15, 25, 35, or 45 mg, NGX267
Received a single dose of either Compound A Form II, designated as A; at each dose, n=3) or matched placebo (n=10). Ten individuals reached the maximum tolerated dose (35 mg), eight subjects (80%) reported a total of 31 adverse events, and two subjects (20%) reported no adverse events. Treatment-emergent adverse events reported in more than one subject treated at this dose were: hypersalivation (4 subjects, 40%), hyperhidrosis (4 subjects, 40%), cold sweats (4 subjects, 40%),
4 subjects, 40%), abdominal discomfort (2 subjects, 20%), and dysgeusia (2 subjects, 20%). Healthy elderly subjects of both sexes (ages 65-80) were randomized in a separate Phase I study.
In a phased study (20 received NGX267 and 6 received placebo), the maximum tolerated oral dose was determined to be 20 mg.

In a randomized, double-blind, placebo-controlled, repeated-dose cohort study of 60 healthy male volunteers (ages 18-54), 48 received Compound A Form II (NGX267; 10, 20,
30, 35 mg once daily for 4 days) and 12 received placebo. NGX267
and its active desmethyl metabolite (NGX292) increased in a dose-proportional manner. Steady state was reached on day 3 of dosing. The apparent elimination half-life of NGX267 was similar across dose levels. Estimated mean values for t1/2 ranged from 7.06 to 7.57 hours on day 0 and 6.58 to 7.14 hours on day 3. Estimated mean values of CL/F ranged from 299.9 to 342.9 ml/min on day 0 and 335.9 ml/min on day 3.
It was 4-373.5 ml/min. The mean percentage of administered dose of NGX267 (as NGX267 or NGX292) recovered in the urine 24 hours after dosing ranged from 0.4001 to 0.4605 across all dose levels.

Example 33 Safety and Preliminary Efficacy of Form III in Sjögren's Syndrome The tolerability, safety, and efficacy of Compound A Form III (NGX267) at single doses of 10 mg, 15 mg, and 20 mg (as capsules according to Example 30) were evaluated compared to placebo. A total of 26 patients were recruited and randomized into 4 treatment groups for 4 treatment periods at 3 study centers. All completed the study and were used for analysis. On each study day, whole-mouth saliva flow velocity was measured as the primary parameter. As a second study parameter, subjective measures of salivary gland dysfunction were measured at doses 2, 4, 6, 12, 14, and 24 using an 8-item visual analog scale.
evaluated after hours. As an additional exploratory parameter, the standard Schirmer test was performed bilaterally at baseline and at 2, 12, 14, and 24 hours after dosing to assess tear production.

All three NGX267 doses were safe, well tolerated and effective with respect to dry mouth. Saliva production and maximal salivary flow 6-24 hours after dosing were significantly greater than those observed with placebo treatment after administration of all three doses of NGX267, with the first 6 and first 24 hours The relationship between dose and saliva production was linear. Subjective measurements were:
There was a significant improvement compared to placebo. The 15 mg dose significantly increased tear production over placebo, but there was no overall treatment effect for this exploratory parameter.

Throughout this description and claims, terms defined when introduced retain their definitions throughout this description and claims.

The invention illustratively described herein suitably is practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. can do. Thus, for example, the terms "comprising", "including"
, “containing,” etc. shall be understood as expansive and non-limiting. In addition, the terms and expressions used herein are used as terms of description, rather than terms of limitation, and the use of such terms and expressions does not represent the meaning of illustration or description. It is not intended to exclude all equivalents of certain features or portions thereof, but rather it is recognized that various modifications are possible within the scope of the invention as claimed. Thus, while the invention has been specifically disclosed with reference to preferred embodiments and optional features, those skilled in the art will appreciate modifications and variations of the invention disclosed herein and embodied therein. , and such modifications and variations are considered to be within the scope of the present invention.

Other embodiments are within the following claims.

Claims (15)

formula:

A crystalline polymorph of Compound A of
The polymorph is
Monohydrate Form III,
(i) the following 2-theta values (±0.2) measured using CuK _α radiation: 12.3;
containing at least one of 17.3, 17.5, 19.9, 21.6, 24,6, 26,3, and 35.4 and having a 2-theta value ranging from 10.8 to 11.9 having an X-ray powder diffraction pattern substantially free of peaks having
(ii) ¹³ C solid-state NMR of the crystalline form, expressed in ppm relative to TMS, with the following chemical shift values: 67.56, 54.60, 47.07, 41.49, 30.70; and at least one resonance with one of 13.77;
(iii) the crystalline form has a ¹³ C solid state NMR of 107.3, 120.3, 127.9;
, 133.4, 144.2, or 161.1, containing the chemical shift difference between the resonance with the largest chemical shift and another resonance;
(iv) the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1039;
1353, 1369, 1369, 1388, 2918, 2974, and 3088 cm ⁻
containing at least one absorption peak having a value selected from ¹ ;
(v) the crystalline form is between 58° C. and 94° C., as determined by differential scanning calorimetry (DSC);
a morphology that exhibits a very broad endothermic peak at 133.7° C. and an endothermic peak with a peak at 134.9° C.;
Anhydrous Form II,
(i) the following single crystal X-ray data: P2(1) a=8.1416(13), (α=90
°), b = 7.9811 (12) (β = 90.761 (2) °, c = 17.878 (3)
, (γ=90°), Å, T=173(1) K, showing a single crystal X-ray characterized by
(ii) the following 2-theta values (±0.2) measured using CuK _α radiation: 9.9;
10.8, 11.8, 11.9, 14.8, 16.2, 18.2, 18.5, 19.8,
having an X-ray powder diffraction pattern containing at least one of 21.3, 22.4, 23.9, 29.2, 29.7, and 33.1;
(iii) the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1906
, 1340, 1447, 2869, 2901, 2951, and 3006-3012 cm
containing at least one absorption peak having a value selected from ^-1 ;
(iv) ¹³ C solid state NMR of the crystalline form with the following chemical shift values expressed in ppm relative to TMS: 175.0; 65.3, 64.0; 45.8, 45.0; 49.3
, 43.6; 39.5.38.8; 28.9, 26.0; 15.4, 14.8;
(v) the crystalline form has a ¹³ C solid state NMR of 109.7 or 111; 129.2 or 130.0; 122.7; 125.7; 131.4;
46.1 or 149.0; and 159.6 or 160.2, and/or Starting at 134.2°C and 135.4°C as measured by DSC
A form having an endothermic peak with a peak at ±0.2°C, substantially no endothermic peak at 106°C to 110°C, and lacking an endothermic peak in the range from about 50°C to about 120°C ;
Monohydrate Form I,
(i) the following 2-theta values (±0.2) measured using CuK _α radiation: 8.8, 1
2.3, 17.5, 19.9, 21.6 23.5, 24.5, 26.3, 28.8, 3
1.6, but the following 2-theta values: 17.3, 17.9, 21.
10.8 lacking at least one of 9, 24.9, 29.3, 30.8, and 33.4
having an X-ray powder diffraction pattern substantially free of peaks with 2-theta values in the range of ~11.9;
(ii) the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1352;
1369, and 1387 cm ⁻¹ , and/or (iii) the crystalline form, as measured by differential scanning calorimetry (DSC), is 107.
showing endothermic peaks at 1°C (starting at 104.85°C) and ^136.17 °C (starting at 133.41°C); contains at least one resonance having one of the following chemical shift values expressed in ppm: 67.09, 54.08, 46.59, 40.97, 30.15, and 13.27; and or (v) the crystalline form has a ¹³ C solid-state NMR of 107.3, 120.3, 127.8, 1
A crystalline polymorph selected from the group consisting of: 33.4, 144.2, or 161.1, containing a chemical shift difference between the resonance with the largest chemical shift and another resonance. 2. The crystalline polymorph of claim 1, which is polymorphic Form II. (a) The crystalline form has the following data: Volume = 1161.6(3) Å3, Z = 4, F(0
00) = 464, theoretical density, Dc = 1.226 Mg/m ³ , absorption coefficient, μ = 0.251 m
further characterized by m ⁻¹ ,
(b) X-ray powder diffraction patterns are measured using _CuKα radiation with the following 2-theta values:
9.9, 10.8, 11.8, 11.9, 14.8, 16.2, 18.2, 18.5, 1
9, 8, 21.3, 22.4, 23.9, 29.2 and 29.7, 33.0, 33.
containing at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all of 1;
(c) an X-ray powder diffraction pattern at a diffraction angle 2θ selected from the group consisting of 14.8° and 19.8° and/or selected from the group consisting of 18.2° and 18.5°; comprising at least one additional peak;
(d) the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 906, 134;
containing at least two, three, four, five, six, or all absorption peaks having values selected from 0, 1447, 2869, 2901, 2951, and 3006-3012 cm ⁻¹ ;
(e) ¹³ C solid state NMR of the crystalline form with the following chemical shift values expressed in ppm relative to TMS: 175.0; 65.3, 64.0; 45.8, 45.0; 49.3, 43
. 6, 39.5; 38.8; 28.9, 26.0; 15.4, 14.8 with at least two, three, four, five, six, seven, or all resonances contains
(f) an X-ray powder diffraction pattern with the following 2-theta values: 9.9, 10.8, 18.5, 1
9.8±0.2 and the ¹³ C solid-state NMR of the crystalline form expressed in ppm relative to TMS with the following chemical shift values: 14.8, 15.4, 26.0, 28.9, 64.0,
^{and /} or (g) the polymorphic form is determined by thermogravimetric analysis (
exhibit less than 1% weight loss as determined by TGA);
3. The crystalline polymorphic Form II of claim 2. 2. The crystalline polymorph of claim 1, which is polymorphic Form III. (a) X-ray powder diffraction patterns are measured using _CuKα radiation with the following 2-theta values:
containing at least two, three, or all of 8.8, 12.30, 17.30, 17.50, 17.80, and 23.0;
(b) X-ray powder diffraction patterns are measured using _CuKα radiation with the following 2-theta values:
12.3 also containing at least one of 19.9.21.6, 24.5, 26.3, and 31.6;
(c) the X-ray powder diffraction pattern is substantially free of peaks with 2-theta values in the range of 10.8 to 11.8;
(d) X-ray powder diffraction pattern measured using _CuKα radiation with the following 2-theta values:
12.2, 17.3, 19.9, 21.6, 24.5, 26.3, and 31.6, and wherein said X-ray powder diffraction pattern is between 10.8 and 11.6. substantially free of peaks with 2-theta values in the range of 8;
(e) X-ray powder diffraction pattern measured using _CuKα radiation with the following 2-theta values:
12.2, 17.3, 17.5, 19.9, 21.6, 24.5, 26.3, and 31
. containing at least two, three, four, five, or all of 2;
(f) ¹³ C solid state NMR of the crystalline form with the following chemical shift values expressed in ppm relative to TMS: 67.56, 54.60, 41.49, 30.70, and 13.77. containing resonances with at least two, three, four, or all of
(g) ¹³ C solid state NMR of 107.3, 120.3, 133.4, 14 for the crystalline form;
4.2, and 161.1, and/
or (h) the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1039, 13
containing at least two, three, four, five, six, or all absorption peaks having values selected from 53, 1369 1369, 1388, 2918, 2974, and 3088 cm ⁻¹ ;
5. Crystalline polymorph Form III according to claim 4. ¹³ C solid-state NMR of the crystalline form with an X-ray powder diffraction pattern containing the following 2-theta values: 12.3, 17.3, 17.5, 19.9, 21.6±0.2 contains resonances with the following chemical shift values expressed in ppm relative to TMS: 13.6, 30.6, 67.5, and the ZnSe ATR-FT-IR absorption spectrum of the crystalline form is 1353, 1
A ^compound ( S)-2-ethyl-8-methyl-1-thia-4,8-diazaspiro [4
. 5] A crystalline monohydrate form of decan-3-one. A process for preparing a crystalline polymorphic form of Compound A according to any one of claims 1 to 6, comprising:

(a) dissolving Compound A in a suitable solvent;
(b) optionally cooling the resulting solution;
(c) waiting a sufficient time to allow the crystalline form to crystallize until Form II crystals precipitate; and (d) filtering said crystalline form;
(i) the crystalline form is polymorphic Form II according to claim 2 or 3 and the solvent is selected from the group consisting of acetone, acetonitrile, cyclohexane, hexane, dioxane and mixed solvents of ethanol and acetonitrile;
(ii) the crystalline form is polymorphic Form III according to any one of claims 4 to 6, obtained by adding 1.3 moles of deionized water to an acetone solution of compound A, Preferably, polymorphic Form III is obtained by reslurrying Compound A and/or polymorphic Form II in deionized water and filtering,
(iii) the crystalline form is polymorphic form I according to claim 1, water-miscible organic solvents containing trace amounts of water (ethanol, ethyl acetate, isopropanol, tert-butyl methyl ether, tetrahydrofuran), water or by slow evaporation of a solution of the compound dissolved in either water or ethyl acetate, or (iv) the crystalline form is a mixture of crystalline polymorphic forms I and II and the solvent is selected from the group consisting of toluene, dichloromethane, 1-butanol, or diethyl ether. A process for converting polymorphic form II to polymorphic form III, wherein the crystalline polymorphic form II according to claim 2 or 3 is dissolved at room temperature and a relative humidity of at least 95% at a temperature of claim 4
7. A process comprising maintaining for a time sufficient to convert to crystalline polymorphic Form III according to any one of paragraphs 1-6. A process for converting polymorphic form I to polymorphic form II according to claim 2 or 3, comprising:
(a) maintaining crystalline Form I at an elevated temperature below the melting point of the crystalline form for a sufficient time to convert the crystalline form to said crystalline polymorphic Form II;
(b) suspending crystalline polymorphic Form I in a solvent selected from the group consisting of acetonitrile, cyclohexane, hexane, dioxane, and mixed solvents of ethanol and acetonitrile, and waiting for a sufficient period of time to obtain the and (c) heating the crystalline polymorphic Form I to a temperature above its melting point to form a molten mass and melting the crystalline form. A process that includes one of: cooling the mass. The resulting crystalline polymorphic Form I, II, or III is measured by its X-ray powder diffraction pattern, ZnSe ATR-FT-IR absorption spectrum DSC in 2-theta values measured using CuK _alpha radiation. A process according to any one of claims 7 to 9, selected by identifying polymorphs by endothermic peaks, TGA and/or resonances measured by ¹³ C solid-state NMR. 4. A process for stably maintaining the crystalline polymorph Form II of claim 2 or 3, comprising maintaining said crystals at room temperature under a dry atmosphere. A pharmaceutical composition comprising a crystalline polymorph according to any one of claims 1 to 6 and at least one pharmaceutically acceptable excipient or carrier, preferably polymorphic form II or I
A pharmaceutical composition wherein II is present in the formulation in an amount of 1 mg to 100 mg, preferably 10 mg to 50 mg, preferably the formulation is granulated. A process for preparing a medicament based formulation of a crystalline polymorph of Compound A, comprising:
(a) Polymorph Form II according to claim 2 or 3, wherein the crystalline polymorph is suitable for oral administration.
and the formulation is directly compressed into a tablet, or (b) the crystalline polymorph is suitable for oral administration. III, which is mixed with one or more excipients (pregelatinized starch, microcrystalline cellulose, colloidal silicon dioxide, and stearic acid), the mixture comprising 1
Process filled into size 4 white opaque hard gelatin bisected capsules that can be used as an oral formulation for immediate release in the gastrointestinal tract to provide 5 mg or 10 mg of polymorphic Form III per capsule. A crystalline polymorphic form according to any one of claims 1 to 6 for use in the treatment of a medical condition responsive to treatment, amelioration or prophylaxis with a muscarinic receptor agonist, wherein Preferably, the daily dose is about 10 mg and 50 mg and/or the condition is a disease or condition associated with cholinergic dysfunction, a disease or condition in which cholinergic function is imbalanced, an acetylcholine receptor Crystalline polymorphic forms, including diseases or conditions associated with impaired activity and diseases or conditions associated with impaired activity of the M1 receptor.
Such diseases and conditions include, but are not limited to, Alzheimer's disease-type senile dementia; Alzheimer's disease (AD); Lewy body dementia combined with Alzheimer's disease and Parkinson's disease; Parkinson's disease; Atrophy; multi-infarct dementia (MID)
, frontotemporal dementia; vascular dementia; stroke/ischemia, MI with stroke/ischemia/head trauma
D; mixed MID and AD; human head trauma; traumatic brain injury; age-related memory impairment; transient global amnesia syndrome; hallucinatory delusional states, affective attention disorder; sleep disturbance; postoperative delirium; adverse effects of tricyclic antidepressants, schizophrenia and Parkinson's disease Adverse effects of certain drugs used; xerostomia, name aphasia, memory loss and/or confusion; psychoses; schizophrenia, schizophrenia with AD, tardive schizophrenia, paraphreny, schizophreniform disorders; anxiety, bipolar disorder, mania; mood stabilization; cognitive impairment after removal of certain gliomas;
Primary age-related tauopathy; chronic traumatic encephalopathy; Pick's disease; progressive supranuclear palsy; corticobasal degeneration), tardive dyskinesia; post-encephalitis amnesia syndrome; sepsis-associated encephalopathy; sepsis-induced delirium;
AIDS-related dementia; memory impairment in autoimmune diseases including lupus, multiple sclerosis, Sjögren's syndrome, chronic fatigue syndrome, and fibromyalgia, splenomegaly, memory impairment in atypical depression or schizophrenia; chemotherapy-induced Cognitive deficits; alcoholic dementia, post-bypass and transplant cognitive deficits, hypothyroidism-related dementia, autism-related cognitive impairment, Down syndrome,
Cognitive impairment due to substance abuse or withdrawal, including nicotine, cannabis, amphetamines, cocaine, attention deficit hyperactivity disorder (ADHD), pain, rheumatism, arthritis, and terminal illness; xerophthalmia, vaginal dryness, dry skin; immune function Disorders; neuroclinic disorders and dysregulation of food intake, including bulimia and anorexia; obesity; congenital ornithine transcarbamylase deficiency; olivopontocerebellar atrophy; alcohol withdrawal; , Huntington's disease; progressive supranuclear palsy; Pick's disease; Friedreich's ataxia; Gilde La Tourette's disease; Down's syndrome; secretions, and dysfunction of gastrointestinal motility and function such as ulcers; urge incontinence, asthma, COPD; brain, nervous system, cardiovascular system, immune system, neuroclinic system, gastrointestinal system, or endocrine and Cerebral amyloid-mediated disorders of one or more of the exocrine glands, eyes, corneas, lungs, prostate, or other organs whose cholinergic function is mediated by muscarinic receptor subtypes; glycogen synthesis; enzyme kinase (GSK3beta)-mediated disorders; tau protein hyperphosphorylation-mediated injury, dysfunction or disease; CNS and PNS hypercholesterolemia and/or hyperlipidemia-mediated injury, dysfunction or disease; hyperglycemia; diabetes; central or peripheral nervous system disease states due to endogenous growth factor-mediated disease or dysfunction involving a combination of additional risk factors; or apolipoproteins disease states involving E; or senile dementia of the Alzheimer's type, Alzheimer's disease (AD), delayed onset of AD symptoms in patients at risk of developing AD, dementia with Lewy bodies, Lewy body disease, cerebral amyloid Angiopathy (CAA), cerebral amyloidosis, frontotemporal dementia, vascular dementia, hyperlipidemia, hypercholesterolemia, multiple infarct dementia (MID), stroke ischemia, stroke/ischemia/
MID with head trauma, mixed MID and Alzheimer's disease, human head trauma, age-related memory impairment, mild cognitive impairment (MCI), MCI leading to AD, bipolar disorder, mania, schizophrenia, apathetic Cholinergic dysfunction has been implicated, including sexual schizophrenia, paraphreny, immune dysfunction, neuroclinic disorders, and dysregulation of food intake, including hyperphagia and anorexia, weight management, obesity, and inflammation. of particular interest in advocating immunotherapy for inflammatory disorders. Preferably, cholinesterase inhibitors, nicotinic agonists, cholinergic precursors and cholinergic enhancers, nootropics, peripheral antimuscarinic agents, M2 muscarinic antagonists, M4 antagonists, benzodiazepine inverse agonists, sigma-1 agonists, Antidepressants, tricyclic antidepressants or antimuscarinic agents, antipsychotics and antischizophrenias, glutamate antagonists and modulators, metabotropics used in the treatment of Parkinson's disease (PD) or depression Glutamate receptor agonist, N
MDA antagonists, AMPA agonists, acetyl-L-carnitine, MAO-B inhibitors, peptides and growth factors, cholesterol lowering agents, antioxidants, GSK-3 beta inhibitors, Wnt-ligands, PKC activators, beta or activators of enzymes involved in the degradation of beta-amyloid, such as gamma-secretase inhibitors, beta-amyloid-degrading agents, neprilysin, insulin-degrading enzymes, or activators of endothelin-converting enzymes;
Beta Amyloid Antiaggregants, Chelating Agents, Beta Amyloid, Antibodies and Immunotherapy Compounds Against Tau Protein Pathology and/or Alpha-Synuclein Pathology, Amyloid Binding Compounds, Cyclooxygenase (COX)-2 Inhibitors, Nonsteroidal Anti-Inflammatories 13. The method of claim 12, further comprising at least one additional pharmacologically active compound selected from the group consisting of drugs, estrogenic agents, estrogen receptor modulators, steroidal neuroprotective agents, and spin trap pharmaceuticals. pharmaceutical composition.

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